Team:NYMU-Taipei/Modeling/MainParts

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Latest revision as of 02:40, 29 October 2013

National Yang Ming University

1 Main parts

Main parts

Backgrounds

Circuit regulators

• ptet regulated promoter (ptet) : when ptet exists, it will bind to ptet regulated promoter (ptet) and represses the promoter.

• pLux/cI hybrid promoter (plux/cI) : when luxR/AHL exists, it will open pLux/cI hybrid promoter (plux/cI), while cI will repress the promoter. What’s more, plux/cI is cI-dominant, which means when cI exists, the hybrid promoter will be repressed whether luxR/AHL exists or not.

Circuit regulation:

• Circuit condition without Nosema:

Without Nosema ceranae, the first circuit will not open, and thus, no ptet will be produced. Since there is no ptet binding to ptet, ptet will not be repressed, which means cI will be produced. After that, cI will bind to pLux/cI hybrid promoter, leading the third circuit to be off.

• Circuit condition with Nosema:

When Nosema ceranae infects the bee, bee’s immune system will be activated, leading to the concentration of ROS (reactive oxygen species) to be high. In response, Bee. coli will enhance the production of oxyR, which forms a complex with ROS and binds to transcription binding site ahead of AhpCp (an oxyR-activated promoter; namely, the sensor), leading to the first circuit to be on.

Since the first circuit is on, the downstream ptet gene will be produced and bind to ptet, leading to the second circuit to be off.

After the second circuit is closed, no cI is produced, leading to the third circuit to be on (no repressor at all). And then, kill protein will be produced and kill Nosema Ceranae.

• Positive feedback:

Besides the LuxI and LuxR(which then form a complex called luxR/AHL) produced by first circuit, the third circuit will also produce LuxI and LuxR, which acts as a positive feedback and strongly enhances the production of killer protein.

The picture shows how LuxI will transfer to AHL, which then bind to LuxR to form a dimer:

• Circuit condition when Nosema is killed:

After Nosema is killed, the sensor will be off and no ptet will be produced.

Without ptet, the ptet will not be repressed, and the second circuit will be on, leading to the production of cI. After cI binds to pLux/cI hybrid promoter, the third circuit to be off. After that, the overall circuit will switch to the beginning stage when there is no Nosema.

Objectives

1. To know how much time it needs from sensing to killing the Nosema after infection

2. To know whether the pathway is effective or not

3. To know the range of kill protein concentration (from the minimal concentration which Is effective to kill Nosema to the maximal concentration which will not do harm to the bees)

System

It is assumed that AHL is abundant and thus the formation of AHL2LuxR complex is determined only by the concentration of LuxR; CI’s mechanism of binding to the promoter is assumed to act as hill effect form; kill protein will sustain a period of time before it is degraded naturally without any other types of decomposition.

Equation1

\frac{d[mRNA_C_I]}{dt}= \frac{1-[ptet]^nptet}{ Kdptet^nptet+ [ptet]^nptet} \times PoPSconstitutive\times\frac{N}{V} -KdegmRNA\times [mRNA_C_I]

Kdptet = dissociation constant of CI

nptet = Hill coefficient of ptet

PoPSptet = promoter strength of ptet

kdegmRNA = degrading constant of sensor promoter mRNA

N = number of plasmid in a single cell

V = volume of a cell

The aim of the equation is to know mRNA_C_I production rate and when it can reach the level to translate the desired concentration.

Results

Figure1: This picture shows kill protein concentration to time under pLux/CI hybrid promoter’s control and LuxR, LuxI’s positive feedback. The result indicates that kill protein concentration will reach its effective level and kill Nosema in time (36 hours after Nosema infection)

Figure2: This picture is a control group model. It shows that without Nosema infection (ROS concentration=0), ethanol production is low enough to be ignored.

Figure3: This is the picture of kill protein concentration to time under promoter CI’s control only(control group)without pLux/CI's regulation.The result shows that kill protein's full expression is too low to kill Nosema.

Discussion

Comparing to the ROS-level control group where ROS remains low when there’s no Nosema infection, kill protein production is highly boasted upon sensing Nosema.

The result indicates that kill protein concentration will reach its effective level and kill Nosema in time (36 hours after sensing Nosema infection, within the 5-day deadline when Nosema spores proliferates and the bee become incurable and infectious to other bees), at the given ROS changes and the enhanced OxyR expression (to learn more about our sensor model, click here).

Circuit design and improvement

Upon using pCI to negatively regulate kill protein production, results show that kill protein's full expression while no CI present is too low to kill Nosema. To solve this problem we subsidized CI promoter into pLux/CI promoter for additional positive feedback to boast kill protein’s production. As the modeling results regarding the modified circuit shown, the consequences are positive. (click here to learn more about our circuit)

Parameters:

Model	Parameter	Description	Value	Unit	Reference
Sensor	KdegOxyR	OxyR degrading rate	10⁷	M^-1 x min^-1	Regulation of the OxyR transcription factor by hydrogen peroxide and the cellular thiol—disulfide status
	KdOxyR*	Dissociation constant of OxyR*	10^-7.33	M
	KOxyR	OxyR producing rate constant	5.012 x 10¹⁴	X
	nOxyR*	Hill coefficient of OxyR*	Ranging from 0.75~3.5,depends on what kind of ROS it react with	X	OxyR: A Molecular Code for Redox-Related Signaling
	N	Copy number	887(cells containing 25％ plasmid bearing cells)	Single piece	Escherichia coli Plasmid Copy Number Assay Improved determination of plasmid copy number using quantitative real-time PCR for monitoring fermentation processes
Ethanol	pyruvate	Initial concentration of pyruvate in MG1655	1.18 x 2	g/L	Expression of pyruvate carboxylase enhances succinate production in Escherichia coli without affecting glucose uptake
	nPDC	Hill coefficient of PDC	2.1	-	Purification, characterization and cDNA sequencing of pyruvate decarboxylase Zygosaccharomyces biporus
	nADH	Hill coefficient of ADH	at high pH values, 30◦C, n=1; at low temperature, n=3	-	Evidence for co-operativity in coenzyme binding to tetrameric Sulfolobus solfataricus alcohol dehydrogenase and its structural basis: ﬂuorescence, kinetic and structural studies of the wild-type enzyme and non-co-operative N249Y mutant
SEIR(exponential)	b	Infection rate constant of Nosema ceranae to the suspected	24/75	Period(days)-1
	r1	Infection rate constant of K12 to the suspected	3/20	Period(days)-1	1. Environment protection administration executive yuan of R.O.C Medical bacteriology of J.A.T
	r2	Infection rate constant of K12 to the latent	3/20	Period(days)-1	2. Environment protection administration executive yuan of R.O.C. 3. Medical bacteriology of J.A.T
	e	rate of the latent turns infectious	1/4	Period(days)-1
	u	Death rate of the infected	1/8	Period(days)-1
	k	Rate of intaking capsule	24/11	Period(days)-1
SEIR(exponential&linear)	S	x(1)	Amount of total population
	E	x(2)	Amount of suspected individuals
	I	x(3)	Amount of individuals in the latent period
	R	x(4)	Amount of infected individuals

Retrieved from "http://2013.igem.org/Team:NYMU-Taipei/Modeling/MainParts"

@@ Line 1: / Line 1: @@
 {{:Team:NYMU-Taipei/Header}}
-=main part=
+=Main parts=
-==Function of the parts:==
+==Backgrounds==
-circuit regulators:
+'''Circuit regulators'''
-• LacI regulated promoter(pLac) : when LacI exists, it will bind to LacI regulated promoter(pLac) and represses the promoter.
+• ptet regulated promoter (ptet) : when ptet exists, it will bind to ptet regulated promoter (ptet) and represses the promoter.
-• pLux/cI hybrid promoter(plux/cI) : when luxR/AHL exists, it will open pLux/cI hybrid promoter(plux/cI), while cI will repress the promoter. What’s more, plux/cI is cI-dominant, which means when cI exists, the hybrid promoter will be repressed whether luxR/AHL exists or not.
+• pLux/cI hybrid promoter (plux/cI) : when luxR/AHL exists, it will open pLux/cI hybrid promoter (plux/cI), while cI will repress the promoter. What’s more, plux/cI is cI-dominant, which means when cI exists, the hybrid promoter will be repressed whether luxR/AHL exists or not.
-Circuit regulation:
+'''Circuit regulation:'''
-• Circuit condition without Nosema:
+• Circuit condition without ''Nosema'':
 [[Image:NYMU_ Circuit condition without Nosema.png|center]]
-Without Nosema Ceranae, the first circuit will not open, and thus, no LacI will be produced. Since there is no LacI binding to pLac, pLac will not be repressed, which means cI will be produced. After that, cI will bind to pLux/cI hybrid promoter, leading the third circuit to be off.
+Without ''Nosema ceranae'', the first circuit will not open, and thus, no ptet will be produced. Since there is no ptet binding to ptet, ptet will not be repressed, which means cI will be produced. After that, cI will bind to pLux/cI hybrid promoter, leading the third circuit to be off.
-• Circuit condition with Nosema:
+• Circuit condition with'' Nosema'':
 [[Image:NYMU_ Circuit condition with Nosema.png|center]]
-When Nosema Ceranae infects the bee, bee’s immune system will be activated, leading to the concentration of ROS (reactive oxygen species) to be high. In response,  Bee. coli will enhance the production of oxyR, which forms a complex with ROS and binds to transcription binding site ahead of trxC (an oxyR-activated promoter; namely, the sensor), leading to the first circuit to be on.
+When ''Nosema ceranae'' infects the bee, bee’s immune system will be activated, leading to the concentration of ROS (reactive oxygen species) to be high. In response,  Bee. coli will enhance the production of oxyR, which forms a complex with ROS and binds to transcription binding site ahead of AhpCp (an oxyR-activated promoter; namely, the sensor), leading to the first circuit to be on.
-Since the first circuit is on, the downstream LacI gene will be produced and bind to pLac, leading to the second circuit to be off.
+Since the first circuit is on, the downstream ptet gene will be produced and bind to ptet, leading to the second circuit to be off.
-After the second circuit is closed, no cI is produced, leading to the third circuit to be on (no repressor at all). And then, kill protein will be produced and kill Nosema Ceranae.
+After the second circuit is closed, no cI is produced, leading to the third circuit to be on (no repressor at all). And then, kill protein will be produced and kill ''Nosema Ceranae''.
-•Positive feedback:
+• Positive feedback:
 [[Image:NYMU_positive feedback.png|center]]
 Besides the LuxI and LuxR(which then form a complex called luxR/AHL) produced by first circuit, the third circuit will also produce LuxI and LuxR, which acts as a positive feedback and strongly enhances the production of killer protein.
-•Circuit condition when Nosema is killed:
+The picture shows how LuxI will transfer to AHL, which then bind to LuxR to form a dimer:
+[[Image:NYMU_luxr ahl dimer.png|center]]
+• Circuit condition when ''Nosema'' is killed:
 [[Image:NYMU_ Circuit condition when Nosema is killed.png|center]]
-After Nosema is killed, the sensor will be off and no lacI will be produced.
+After ''Nosema'' is killed, the sensor will be off and no ptet will be produced.
-Without LacI, the pLac will not be repressed, and the second circuit will be on, leading to the production of cI. After cI binds to pLux/cI hybrid promoter, the third circuit to be off. After that, the overall circuit will switch to the beginning stage when there is no Nosema.
+Without ptet, the ptet will not be repressed, and the second circuit will be on, leading to the production of cI. After cI binds to pLux/cI hybrid promoter, the third circuit to be off. After that, the overall circuit will switch to the beginning stage when there is no'' Nosema''.
-==Aims==
+==Objectives==
+. To know how much time it needs from sensing to killing the ''Nosema'' after infection
-. To know how much time it needs from sensing to killing the Nosema after infection
 . To know whether the pathway is effective or not
-. To know the range of kill protein concentration (from the minimal concentration which Is effective to kill Nosema to the maximal concentration which will not do harm to the bees)
-It is assumed that AHL is abundant and thus the formation of AHL2LuxR complex is determined only by the concentration of LuxR; CI’s mechanism of binding to the promoter is assumed to act as hill effect form; kill protein will sustain a period of time before it is degraded naturally without any other types of decomposition.
+. To know the range of kill protein concentration (from the minimal concentration which Is effective to kill ''Nosema'' to the maximal concentration which will not do harm to the bees)
-==Equations==
-'''Equation1:'''
+==System==
+It is assumed that AHL is abundant and thus the formation of AHL2LuxR complex is determined only by the concentration of LuxR; CI’s mechanism of binding to the promoter is assumed to act as hill effect form; kill protein will sustain a period of time before it is degraded naturally without any other types of decomposition.
+'''Equation1'''
 <html>
 <div lang="latex" class="equation">
-\frac{d[mRNACI]}{dt}= \frac{1-[LacI]^nLacI}{ KdLacI^nLacI+ [LacI]^nLacI}
+\frac{d[mRNA_C_I]}{dt}= \frac{1-[ptet]^nptet}{ Kdptet^nptet+ [ptet]^nptet}
 \times PoPSconstitutive\times\frac{N}{V}
--KdegmRNA\times [mRNACI]
+-KdegmRNA\times [mRNA_C_I]
 </div>
 </html>
-KdLacI = dissociation constant of CI
+Kdptet = dissociation constant of CI
-nLacI = Hill coefficient of LacI
+nptet = Hill coefficient of ptet
-PoPSpLac = promoter strength of pLac
+PoPSptet = promoter strength of ptet
 kdegmRNA = degrading constant of sensor promoter mRNA
@@ Line 71: / Line 78: @@
 V = volume of a cell
-''' The aim of the equation is to knowmRNACI production rate and when it can reach the level to translate the desired concentration.'''
+''' The aim of the equation is to know mRNA_C_I production rate and when it can reach the level to translate the desired concentration.'''
+[[read more1]]
-'''Equation2:'''
+'''Equation2'''
 <html>
 <div lang="latex" class="equation">
-\frac{d[mRNALacI]}{dt}= \frac{1-[ROSoxyR]^nROSoxyR }{ KdROSoxyR ^nROSoxyR + [ROSoxyR]^nROSoxyR} PoPStrxC\times\frac{N}{V}-KdegmRNA[mRNALacI]
+\frac{d[mRNA_L_a_c_I]}{dt}= \frac{1-[ROSoxyR]^nROSoxyR }{ KdROSoxyR ^nROSoxyR + [ROSoxyR]^nROSoxyR} PoPSAhpCp\times\frac{N}{V}-KdegmRNA[mRNA_L_a_c_I]
 </div>
 </html>
@@ Line 84: / Line 93: @@
 nROSoxyR = Hill coefficient of ROSoxyR
-PoPStrxC = promoter strength of trxC
+PoPSAhpCp = promoter strength of AhpCp
 kdegmRNA = degrading constant of sensor promoter mRNA
@@ Line 92: / Line 101: @@
 V = volume of a cell
-''' The aim of the equation is to know how trxC promoter strength (in PoPS) influences the production of lacI.'''
+''' The aim of the equation is to know how AhpCp promoter strength (in PoPS) influences the production of ptet.'''
+[[read more2]]
 '''Equation3:'''
@@ Line 98: / Line 109: @@
 <html>
 <div lang="latex" class="equation">
-\frac{d[mRNAOxyR]}{dt}=PoPSconstitutive\times\frac{N}{V}-KdegmRNA\times [mRNAOxyR]</div>
+\frac{d[mRNA_O_x_y_R]}{dt}=PoPSconstitutive\times\frac{N}{V}-KdegmRNA\times [mRNAOxyR]</div>
 </html>
@@ Line 111: / Line 122: @@
 ''' The aim of the equation is to know the production rate ofmRNAoxyR, and choose the proper constitutive promoter for boosting OxyR's concentration.'''
+[[read more3]]
 '''Equation4:'''
 <html> <div lang="latex" class="equation">
-\frac{d[mRNALuxI]}{dt}= \frac{1-[ROSoxyR]^n ROSoxyR }{ Kd ROSoxyR ^n ROSoxyR + [ROSoxyR]^n ROSoxyR I} PoPStrxC\times\frac{N}{V}+\frac{1-[ LuxRAHL]^nLuxRAHL}{ KdLuxRAHL ^nLuxRAHL + [LuxRAHL]^nLuxRAHL}\times\frac{1-{1-[CI leak]^nCI}}{ KdCI^nCI+[CI]^nCI}\times PoPSLuxCI\times\frac{N}{V}-KdegmRNA[mRNALuxI]
+\frac{d[mRNA_L_u_x_I]}{dt}= \frac{1-[ROSoxyR]^n ROSoxyR }{ Kd ROSoxyR ^n ROSoxyR + [ROSoxyR]^n ROSoxyR I} PoPSAhpCp\times\frac{N}{V}+\frac{1-[ LuxRAHL]^nLuxRAHL}{ KdLuxRAHL ^nLuxRAHL + [LuxRAHL]^nLuxRAHL}\times\frac{1-{1-[CI leak]^nCI}}{ KdCI^nCI+[CI]^nCI}\times PoPSLuxCI\times\frac{N}{V}-KdegmRNA[mRNALuxI]
 </div> </html>
@@ Line 122: / Line 134: @@
 nROSoxyR = Hill coefficient of ROSoxyR
-PoPStrxC = promoter strength of trxC
+PoPSAhpCp = promoter strength of AhpCp
 KdLuxRAHL = dissociation constant ofLuxRAHL
@@ Line 140: / Line 152: @@
 V = volume of a cell
-''' The aim of the equation is to know how trxC promoter strength (in PoPS) influences the production of LuxI. '''
+''' The aim of the equation is to know how AhpCp promoter strength (in PoPS) influences the production of LuxI. '''
+[[read more4]]
 '''Equation5:'''
 <html> <div lang="latex" class="equation">
-\frac{d[mRNALuxR]}{dt}= \frac{1-[ROSoxyR]^n ROSoxyR }{ Kd ROSoxyR ^n ROSoxyR + [ROSoxyR]^n ROSoxyR I} \times PoPStrxC\times\frac{N}{V}+\frac{1-[ LuxRAHL]^nLuxRAHL}{ KdLuxRAHL ^nLuxRAHL + [LuxRAHL]^nLuxRAHL}\times\frac{1-{1-[CI leak]^nCI}}{ KdCI^nCI+[CI]^nCI}\times PoPSLuxCI\times\frac{N}{V}-KdegmRNA[mRNALuxR]
+\frac{d[mRNA_L_u_x_R]}{dt}= \frac{1-[ROSoxyR]^n ROSoxyR }{ Kd ROSoxyR ^n ROSoxyR + [ROSoxyR]^n ROSoxyR I} \times PoPSAhpCp\times\frac{N}{V}+\frac{1-[ LuxRAHL]^nLuxRAHL}{ KdLuxRAHL ^nLuxRAHL + [LuxRAHL]^nLuxRAHL}\times\frac{1-{1-[CI leak]^nCI}}{ KdCI^nCI+[CI]^nCI}\times PoPSLuxCI\times\frac{N}{V}-KdegmRNA[mRNALuxR]
 </div> </html>
@@ Line 152: / Line 166: @@
 nROSoxyR = Hill coefficient of ROSoxyR
-PoPStrxC = promoter strength of trxC
+PoPSAhpCp = promoter strength of AhpCp
 KdLuxRAHL = dissociation constant ofLuxRAHL
@@ Line 170: / Line 184: @@
 V = volume of a cell
-''' The aim of the equation is to know how trxC promoter strength (in PoPS) influences the production of LuxR. '''
+''' The aim of the equation is to know how AhpCp promoter strength (in PoPS) influences the production of LuxR. '''
+[[read more5]]
-'''Equation6:'''
+'''Equation6'''
 <html> <div lang="latex" class="equation">
-\frac{d[mRNAkill]}{dt}= \frac{1-[ LuxRAHL]^nLuxRAHL}{ KdLuxRAHL ^nLuxRAHL + [LuxRAHL]^nLuxRAHL}\times\frac{1-{1-[CI leak]^nCI}}{ KdCI^nCI+[CI]^nCI}\times PoPSLuxCI\times\frac{N}{V}-KdegmRNA[mRNAkill]
+\frac{d[mRNA_k_i_l_l]}{dt}= \frac{1-[ LuxRAHL]^nLuxRAHL}{ KdLuxRAHL ^nLuxRAHL + [LuxRAHL]^nLuxRAHL}\times\frac{1-{1-[CI leak]^nCI}}{ KdCI^nCI+[CI]^nCI}\times PoPSLuxCI\times\frac{N}{V}-KdegmRNA[mRNA_k_i_l_l]
 </div> </html>
@@ Line 188: / Line 204: @@
 PoPSLuxcI = promoter strength of LuxcI hybrid promoter
-kdegmRNA = degrading constant of sensor promoter Mrna
+kdegmRNA = degrading constant of sensor promoter mRNA
 N = number of plasmid in a single cell
@@ Line 194: / Line 210: @@
 V = volume of a cell
-'''The aim of the equation is to knowmRNA of kill protein production rate and when it can reach the level ofthe desired concentration. '''
+'''The aim of the equation is to know mRNA of kill protein production rate and when it can reach the level of the desired concentration. '''
-'''Equation7:'''
+[[read more6]]
+[[to learn more about how we characterize pLuxCI, click here]]
+'''Equation7'''
 <html> <div lang="latex" class="equation">
-\frac{d[Mrnat7]}{dt}= \frac{1-[ROSoxyR]^n ROSoxyR }{ Kd ROSoxyR ^n ROSoxyR + [ROSoxyR]^nROSoxyR }\timesPoPStrxC\times\frac{N}{V}\times {a}+ \frac{1-[T7]^nT7}{ Kd T7^nT7 + [T7]^nT7}\times{PoPST7}\times\ frac{N}{V}-KdegmRNA\times [mRNAT7]
+\frac{d[mRNA_t_7]}{dt}= \frac{1-[ROSoxyR]^n ROSoxyR }{ Kd ROSoxyR ^n ROSoxyR + [ROSoxyR]^nROSoxyR }\timesPoPSAhpCp\times\frac{N}{V}\times {a}+ \frac{1-[T7]^nT7}{ Kd T7^nT7 + [T7]^nT7}\times{PoPST7}\times\ frac{N}{V}-KdegmRNA\times [mRNA_T_7]
 </div> </html>
@@ Line 206: / Line 227: @@
 nROSoxyR = Hill coefficient of ROSoxyR
-PoPStrxC = promoter strength of trxC
+PoPSAhpCp = promoter strength of AhpCp
 KdT7 = dissociation constant of T7
@@ Line 214: / Line 235: @@
 PoPST7 = promoter strength of T7 promoter
-kdegmRNA = degrading constant of sensor promoter Mrna
+kdegmRNA = degrading constant of sensor promoter mRNA
 N = number of plasmid in a single cell
@@ Line 220: / Line 241: @@
 V = volume of a cell
-'''The aim of the equation is to knowmRNA of T7 polymerase production rate and when it can reach the level to translate enough T7 polymerase. '''
+'''The aim of the equation is to know mRNA of T7 polymerase production rate and when it can reach the level to translate enough T7 polymerase. '''
+[[read more7]]
 '''Equation8:'''
@@ Line 226: / Line 249: @@
 <html>
 <div lang="latex" class="equation">
-\frac{d[mRNAPDC]}{dt}=\frac{ [T7]^{nT7} }{ KdT7^{nT7}+[T7]^{nY7} }\times{PoPST7}\times\frac{N}{V}-KdegmRNA\times[mRNAPDC]
+\frac{d[mRNA_P_D_C]}{dt}=\frac{ [T7]^{nT7} }{ KdT7^{nT7}+[T7]^{nY7} }\times{PoPST7}\times\frac{N}{V}-KdegmRNA\times[mRNAPDC]
 </div>
 </html>
@@ Line 243: / Line 266: @@
 '''The aim of the equation is to knowmRNA of enzyme PDC production rate and when it can reach the level to translate enough PDC. '''
+[[read more8]]
 '''Equation9:'''
@@ Line 248: / Line 273: @@
 <html>
 <div lang="latex" class="equation">
-\frac{d[mRNAADH]}{dt}=\frac{ [T7]^{nT7} }{ KdT7^{nT7}+[T7]^{nY7} }\times{PoPST7}\times\frac{N}{V}-KdegmRNA\times[mRNAADH]
+\frac{d[mRNA_A_D_H]}{dt}=\frac{ [T7]^{nT7} }{ KdT7^{nT7}+[T7]^{nY7} }\times{PoPST7}\times\frac{N}{V}-KdegmRNA\times[mRNAADH]
 </div>
 </html>
@@ Line 266: / Line 291: @@
 '''The aim of the equation is to knowmRNA of enzyme ADH production rate and when it can reach the level to translate enough ADH. '''
-'''Equation10: '''
+[[read more9]]
+'''Equation10 '''
 <html>
 <div lang="latex" class="equation">
-\frac{d[CI]}{dt}={RBS}\times{[mRNACI]} -KdegCI\times{[CI]}
+\frac{d[CI]}{dt}={RBS}\times{[mRNA_C_I]} -KdegCI\times{[CI]}
 </div>
 </html>
@@ Line 280: / Line 308: @@
 '''The aim of the equation is to know the production rate of CI and when it can reach the concentration to repress LuxCI hybrid promoter. '''
-'''Equation11:'''
+[[read more10]]
+'''Equation11'''
 <html>
 <div lang="latex" class="equation">
-\frac{d[LacI]}{dt}={RBS}\times{[mRNA LacI]} -Kdeg LacI \times{[LacI]}
+\frac{d[ptet]}{dt}={RBS}\times{[mRNA_L_a_c_I]} - {Kdeg ptet} \times{[ptet]}
 </div>
 </html>
@@ Line 290: / Line 320: @@
 RBS = binding site strength
-KdegLacI = degrading constant of LacI
+Kdegptet = degrading constant of ptet
-'''The aim of the equation is to knowLacI production rate and when it can reach the concentration to repress promoter LacI.'''
+'''The aim of the equation is to known ptet production rate and when it can reach the concentration to repress promoter ptet.'''
-'''Equation12:''
+[[read more11]]
+'''Equation12''
 <html>
 <div lang="latex" class="equation">
-\frac{d[LuxI]}{dt}={RBS}\times{[mRNA LuxI]} -KAHL\times{[LuxI]}
+\frac{d[LuxI]}{dt}={RBS}\times{[mRNA_L_u_x_I]} -KAHL\times{[LuxI]}
   -Kdeg {[LuxI]} \times{[LuxI]}
 </div>
@@ Line 311: / Line 343: @@
 '''The aim of the equation is to know the production rate of LuxI concerning the LuxIAHL reaction and LuxI degrading. '''
-'''Equation13:''
+[[read more12]]
+'''Equation13''
 <html>
 <div lang="latex" class="equation">
-\frac{d[LuxR]}{dt}={RBS}\times{[mRNA LuxR]} – {KmAHL}\times{[AHL]}
+\frac{d[LuxR]}{dt}={RBS}\times{[mRNA_L_u_x_R]} – {KmAHL}\times{[AHL]}
 -	{Kdeg LuxR} \times{[LuxR]} \times {KmAHL}\{times[AHL]^{2}} \times{[LuxR]}- KdegAHL\times{[AHL]}
 </div>
@@ Line 331: / Line 365: @@
 '''The aim of the equation is to know the production rate of LuxR concerning the 2AHL + LuxR to (AHL)2/LuxR reaction , AHL to LuxI reaction, and LuxR, AHL degrading. '''
+[[read more13]]
 '''Equation14:''
@@ Line 350: / Line 386: @@
 '''The aim of the equation is to know the production rate of AHL concerning the LuxIAHL reaction, 2AHL + LuxR to (AHL)2/LuxR reaction, (AHL)2/LuxR complex to 2AHL + LuxR reaction and AHL degrading. '''
-'''Equation15:''
+[[read more14]]
-<html>
-<div lang="latex" class="equation">
-\frac{d[T7]}{dt}={RBS}\times{[mRNAT7]} –KdegT7\times{[T7]}
-</div>
-</html>
-RBS = binding site strength
-KdegT7 = degrading constant of T7
-'''The aim of the equation is to know the threshold concentration of oxyR to conquer the terminal and when T7 polymerase can reach the required concentration to activate T7 promoter. '''
-'''Equation16:''
-<html>
-<div lang="latex" class="equation">
-\frac{d[PDC]}{dt}=RBS\times{PoPST7effect}\times\frac{N}{V}-{kdegPDC}\times{[PDC]}
-</div>
-</html>
-RBS = binding site strength
-PoPST7 = promoter strength of T7 promoter
-N = number of plasmid in a single cell
-V = volume of a cell
-KdegPDC = degrading constant of PDC (pyruvate decarboxylase)
-'''The aim of the equation is to know PDC production rate and when it can reach the concentration of ethanol pathway equilibrium.'''
-'''Equation17:''
+'''Equation17'''
 <html>
@@ Line 405: / Line 410: @@
 '''The aim of the equation is to know ADH production rate and when it can reach the concentration of ethanol pathway equilibrium.'''
-'''Equation18:''
+[[read more17]]
+'''Equation18'''
 <html>
 <div lang="latex" class="equation">
@@ Line 423: / Line 429: @@
 '''The aim of the equation is to know the production rate of AHL concerning the LuxIAHL reaction, 2AHL + LuxR to (AHL)2/LuxR reaction, (AHL)2/LuxR complex to 2AHL + LuxR reaction and AHL degrading. '''
-'''Equation19:''
+[[read more18]]
+'''Equation19'''
 <html>
 <div lang="latex" class="equation">
-\frac{d[kill]}{dt}={RBS}\times{[mRNAkill]} –Kdegkill\times{[kill]}
+\frac{d[kill]}{dt}={RBS}\times{[mRNA_k_i_l_l]} –Kdegkill\times{[kill]}
 </div>
 </html>
@@ Line 435: / Line 443: @@
 Kdegkill = degrading constant of kill
-'''The aim of the equation is to knowkiller protein production rate and when it can reach the effective concentration to killNosema. '''
+'''The aim of the equation is to knowkiller protein production rate and when it can reach the effective concentration to kill ''Nosema''. '''
-'''Equation20:''
+[[read more19]]
-<html>
-<div lang="latex" class="equation">
-\frac{d[ethanol]}{dt}=Kpyruvateacetaldehyde\times{Kacetaldehydeethanol}\times\frac{[PDC]^{nPDC} }{ KdPDC^{nPDC}+[PDC]^{nPDC}}\times\frac{[ADH]^{nADH} }{ KdADH^{nADH}+[ADH]^{nADH}}-Km\times\frac{[ADH]^{nADH}}{ KdADH^{nADH}+[ADH]^{nADH}}\times{[ethonal]}
-</div>
-</html>
-<br>
-Kpyruvateacetaldehyde = pyruvate→acetaldehyde reaction rate constant
-Kacetaldehydeethanol = acetaldehyde→ethanol reaction rate constant
-Km = ethanol→acetaldehyde reaction rate constant
-KdADH = dissociation constant of ADH
-'''The aim of the equation is to know ethanol production rate and when it can reach the concentration to kill the spores of Nosema Ceranae.''
-==Explanation:==
-<html>
-<div lang="latex" class="equation">
-\frac{d[mRNACI]}{dt}= \frac{1-[LacI]^nLacI}{ KdLacI^nLacI+ [LacI]^nLacI} \times PoPSconstitutive\times\frac{N}{V}-KdegmRNA\times [mRNACI]
-</div>
-</html>
-In this equation, <html><span lang="latex"> PoPSpLac </span></html>
-represents the promoter strength of LacI promoter, which is measured by the rate of RNApolymerase binding to the starting site of DNA transcription; <html><span lang="latex"> \frac{1-[LacI]^nLacI }{ KdLacI^nLacI+[LacI]^nLacI } </span></html>
-represents the hill effect of repressor LacI to LacI promoter. Because LacI is a repressor, the numerator is <html><span lang="latex"> {1-[LacI]^nLacI }</span></html>
-.
-	For the section of the equation, <html><span lang="latex"> \frac{1-[LacI]^nLacI}{ KdLacI^nLacI+ [LacI]^nLacI} \times
-PoPSconstitutive\times\frac{N}{V}</span></html>represents the synthesizing rate of mRNACI under the influence of LacI and LacI promoter; <html><span lang="latex"> -{kdegmRNA}\times {[mRNACI]} </span></html>represents the degrading rate of mRNA
-<html>
-<div lang="latex" class="equation">
-\frac{d[CI]}{dt}={RBS}\times{[mRNACI]} -KdegCI\times[CI]
-</div>
-</html>
-In this equation, RBS represents ribosome binding site strength, which is the affinity of ribosome to the starting site of mRNA.
-	For the section of the equation, <html><span lang="latex"> RBS\times [mRNAcI] </span></html> represents the synthesizing rate of CI; <html><span lang="latex">  -{kdegCI}\times {[CI]} </span></html> represents the degrading rate of CI.
-	Since LuxCI hybrid promoter is CI dominant, the presence of CI block the promoter and kill protein is not produced. This situation happens when there is no Nosema in bees.
-<html>
-<div lang="latex" class="equation">
-\frac{d[mRNALacI]}{dt}= \frac{1-[ROSoxyR]^nROSoxyR }{ KdROSoxyR ^nROSoxyR + [ROSoxyR]^nROSoxyR} \times {PoPStrxC}\times\frac{N}{V}-KdegmRNA[mRNALacI]
-</div>
-</html>
-In this equation, PoPStrxC represents the promoter strength of promotertrxC, which is measured by the rate of RNApolymerase binding to the starting site of DNA transcription; <html><span lang="latex"> \frac{1-[ROSoxyR]^nROSoxyR }{ KdROSoxyR ^nROSoxyR + [ROSoxyR]^nROSoxyR} </span></html> represents the hill effect of activator ROSoxyR to trxCpromoter.
-For the section of the equation, <html><span lang="latex">  \frac{1-[ROSoxyR]^nROSoxyR }{ KdROSoxyR ^nROSoxyR + [ROSoxyR]^nROSoxyR} \times {PoPStrxC}\times\frac{N}{V}</span></html> represents the synthesizing rate of mRNALacI under the influence of activator ROSoxyR complex and trxC promoter; <html><span lang="latex"> -kdegmRNA\times [mRNAlacI] </span></html>represents the degrading rate of mRNALacI.
-<html>
-<div lang="latex" class="equation">
-\frac{d[LacI]}{dt}={RBS}\times{[mRNA LacI]} –{Kdeg LacI} \times{[LacI]}
-</div>
-</html>
-In this equation, RBS represents ribosome binding site strength, which is the affinity of ribosome to the starting site of mRNA.
-For the section of the equation, <html><span lang="latex"> RBS\times [mRNALacI] </span></html> represents the synthesizing rate of LacI; <html><span lang="latex"> -KdegCI\times [LacI] </span></html> represents the degrading rate of LacI .
-LacI will repress pLac promoter, which leads to no production of CI. Without CI, LuxCI hybrid promoter will no longer be repressed, and kill protein can thus be generated. This situation happens when Nosema exists and the sensor promoter is turned on.
-<html>
-<div lang="latex" class="equation">
-\frac{d[mRNAOxyR]}{dt}=PoPSconstitutive\times\frac{N}{V}-KdegmRNA\times [mRNAOxyR]</div>
-</html>
-In this equation, PoPSconstitutive represents the promoter strength of the constitutive promoter J23102, which is measured by the rate of RNApolymerase binding to the starting site of DNA transcription.
-For the section of the equation, <html><span lang="latex"> PoPSconstitutive\times\frac{N}{V}</span></html> represents the synthesizing rate of mRNA"oxyR" ; <html><span lang="latex"> -KdegmRNA\times [mRNAOxyR]</span></html> represents the degrading rate of mRNA"oxyR" .
-<html> <div lang="latex" class="equation">
-\frac{d[mRNALuxI]}{dt}= \frac{1-[ROSoxyR]^n ROSoxyR }{ Kd ROSoxyR ^n ROSoxyR + [ROSoxyR]^n ROSoxyR I} \times PoPStrxC\times\frac{N}{V}+\frac{1-[ LuxRAHL]^nLuxRAHL}{ KdLuxRAHL ^nLuxRAHL + [LuxRAHL]^nLuxRAHL}\times\frac{1-{1-[CI leak]^nCI}}{ KdCI^nCI+[CI]^nCI}\times PoPSLuxCI\times\frac{N}{V}-KdegmRNA[mRNALuxI]
-</div> </html>
-In this equation, PoPStrxC,PoPSLuxcI represents the promoter strength of promoter trxC and LuxcI hybrid promoter, which is measured by the rate of RNApolymerase binding to the starting site of DNA transcription; <html><span lang="latex"> \frac{1-[ROSoxyR]^n ROSoxyR }{ Kd ROSoxyR ^n ROSoxyR + [ROSoxyR]^n ROSoxyR I} </span></html> represents the hill effect of activator ROSoxyR complex to trxCpromoter; <html><span lang="latex">\frac{1-[ LuxRAHL]^nLuxRAHL}{ KdLuxRAHL ^nLuxRAHL + [LuxRAHL]^nLuxRAHL} </span></html>represents the hill effect of activator LuxRAHL to LuxcI hybrid promoter; <html><span lang="latex"> \frac{1-[CI leak]^nCI}{ KdCI^nCI+[CI]^nCI}</span></html> represents the hill effect of repressor CI, while <html><span lang="latex"> frac{1-{1-[CI leak]^nCI}}{ KdCI^nCI+[CI]^nCI}</span></html> takes CI leaking into consideration (promoter pLac will not close thoroughly even though CI exists).
-	For the section of the equation, <html><span lang="latex"> \frac{1-[ROSoxyR]^n ROSoxyR }{ Kd ROSoxyR ^n ROSoxyR + [ROSoxyR]^n ROSoxyR I} \times PoPStrxC\times\frac{N}{V}</span></html> represents the synthesizing rate of mRNALuxI under the influence of activator ROSoxyR complex and trxC promoter; <html><span lang="latex"> \frac{1-[ LuxRAHL]^nLuxRAHL}{ KdLuxRAHL ^nLuxRAHL + [LuxRAHL]^nLuxRAHL}\times\frac{1-{1-[CI leak]^nCI}}{ KdCI^nCI+[CI]^nCI}\times PoPSLuxCI\times\frac{N}{V}</span></html> represents the synthesizing rate of mRNALuxI under the influence of LuxRAHL complex, CI repression and leaking, and LuxcI hybrid promoter; <html><span lang="latex"> -kdegmRNA\times [mRNAluxI]</span></html>represents the degrading rate of mRNAluxI.
-<html>
-<div lang="latex" class="equation">
-\frac{d[LuxI]}{dt}={RBS}\times{[mRNA LuxI]} -KAHL\times{[LuxI]}
- -Kdeg {[LuxI]} \times{[LuxI]}
-</div>
-</html>
-In this equation, RBS represents ribosome binding site strength, which is the affinity of ribosome to the starting site of mRNA; KAHL represents the rate constant of AHL  to LuxI.
-	For the section of the equation, <html><span lang="latex"> {RBS}\times {[mRNALuxI]}</span></html> represents the synthesizing rate of LuxI; KAHL[LuxI] represents the transforming rate of AHL to LuxI; <html><span lang="latex"> -KdegLuxI\times [LuxI] </span></html> represents the degrading rate of LuxI.
-	LacI will repress pLac promoter, which leads to no production of CI. Without CI, LuxCI hybrid promoter will no longer be repressed, and kill protein can thus be generated. This situation happens when Nosema exist, and when the sensor promoter is turned on.
-<html> <div lang="latex" class="equation">
-\frac{d[mRNALuxR]}{dt}= \frac{1-[ROSoxyR]^n ROSoxyR }{ Kd ROSoxyR ^n ROSoxyR + [ROSoxyR]^n ROSoxyR I} \times PoPStrxC\times\frac{N}{V}+\frac{1-[ LuxRAHL]^nLuxRAHL}{ KdLuxRAHL ^nLuxRAHL + [LuxRAHL]^nLuxRAHL}\times\frac{1-{1-[CI leak]^nCI}}{ KdCI^nCI+[CI]^nCI}\times PoPSLuxCI\times\frac{N}{V}-KdegmRNA[mRNALuxR]
-</div> </html>
-In this equation, PoPStrxC,PoPSLuxcI represents the promoter strength of promoter trxC and LuxcI hybrid promoter, which is measured by the rate of RNApolymerase binding to the starting site of DNA transcription; <html><span lang="latex"> \frac{1-[ROSoxyR]^n ROSoxyR }{ Kd ROSoxyR ^n ROSoxyR + [ROSoxyR]^n ROSoxyR I} </span></html> represents the hill effect of activator ROSoxyR complex to trxCpromoter; <html><span lang="latex">\frac{1-[ LuxRAHL]^nLuxRAHL}{ KdLuxRAHL ^nLuxRAHL + [LuxRAHL]^nLuxRAHL} </span></html>represents the hill effect of activator LuxRAHL to LuxcI hybrid promoter; <html><span lang="latex">\ frac{1-[CI leak]^nCI}{ KdCI^nCI+[CI]^nCI}</span></html> represents the hill effect of repressor CI, while <html><span lang="latex"> frac{1-{1-[CI leak]^nCI}}{ KdCI^nCI+[CI]^nCI}</span></html> takes CI leaking into consideration (promoter pLac will not close thoroughly even though CI exists).
-For the section of the equation, <html><span lang="latex"> \frac{1-[ROSoxyR]^n ROSoxyR }{ Kd ROSoxyR ^n ROSoxyR + [ROSoxyR]^n ROSoxyR I} \times PoPStrxC\times\frac{N}{V}</span></html> represents the synthesizing rate of mRNALuxR under the influence of activator ROSoxyR complex and trxC promoter; <html><span lang="latex"> \frac{1-[ LuxRAHL]^nLuxRAHL}{ KdLuxRAHL ^nLuxRAHL + [LuxRAHL]^nLuxRAHL}\times\frac{1-{1-[CI leak]^nCI}}{ KdCI^nCI+[CI]^nCI}\times PoPSLuxCI\times\frac{N}{V}</span></html> represents the synthesizing rate of mRNALuxR under the influence of LuxRAHL complex, CI repression and leaking, and LuxcI hybrid promoter; <html><span lang="latex"> -kdegmRNA\times [mRNAluxI]</span></html>represents the degrading rate of mRNAluxR.
-<html>
-<div lang="latex" class="equation">
-\frac{d[LuxR]}{dt}={RBS}\times{[mRNA LuxR]} – {KmAHL}\times{[AHL]}
--	{Kdeg LuxR} \times{[LuxR]} \times {KmAHL}\{times[AHL]^{2}} \times{[LuxR]}- KdegAHL\times{[AHL]}
-</div>
-</html>
-In this equation, RBS represents ribosome binding site strength, which is the affinity of ribosome to the starting site of mRNA; KmAHL represents the rate constant of AHLLuxI; KonAHL represents the rate constant of 2AHL+LuxR to AHL2LuxR complex.
-For the section of the equation, <html><span lang="latex">{ RBS}\times{ [mRNALuxR]} </span></html> represents the synthesizing rate of LuxR; <html><span lang="latex">{ -KmAHL}\times{ [AHL]} </span></html> represents the transforming rate of AHL to LuxI; <html><span lang="latex">{-KdegLuxR}\times {[LuxR]} </span></html> represents the degrading rate of LuxR; <html><span lang="latex">{ -KonAHL}\times{ [AHL]^2}\times{ [LuxR]}</span></html> represents the transforming rate of 2AHL+LuxR to AHL2LuxR complex; <html><span lang="latex">{-KdegLuxI}\times {[AHL]} </span></html>
-represents the degrading rate of AHL.
-<html>
-<div lang="latex" class="equation">
-\frac{d[AHL]}{dt}={KAHL}\times{[LuxI]}+2\times KoffAHL \times [AHLLuxR] - 2\times KonAHL \times {[AHL]}^{2}\times {[LuxR]} - KdegAHL\times{[AHL]}
-</div>
-</html>
-In this equation, KAHL represents the rate constant of AHL to LuxI; KoffAHL represents the rate constant of AHL2LuxR complex to 2AHL+ LuxR, while KonAHL represents the reverse rate constant.
-For the section of the equation, <html><span lang="latex"> {KAHL}\times {[LuxI]} </span></html> represents the transforming rate of LuxI to AHL; <html><span lang="latex"> {2}\times {KoffAHL}\times{ [AHLLuxR]} </span></html> represents the transforming rate of AHL2LuxR complex to AHL and LuxR, the constant 2 means that each complex turns into two AHL; <html><span lang="latex"> -{2}\times {KonAHL}\times {[AHL]^2}\times {[LuxR]} </span></html> represents the transforming rate of 2AHL+LuxR to AHL2LuxR complex; <html><span lang="latex"> {-KdegLuxI}\times {[LuxI]}</span></html> represents the degrading rate of LuxI.
-Since LuxI will transform to AHL, and 2AHL will combine to LuxR to form AHL2LuxR complex. Because AHL2LuxR will then bind to LuxCI hybrid promoter and activate kill gene, LuxR gene, and LuxI gene. In this way, kill protein will kill Nosema, while LuxR, LuxI will trigger the positive feedback and enhance the production of kill protein.
-<html>
-<div lang="latex" class="equation">
-\frac{d[kill]}{dt}={RBS}\times{[mRNAkill]} –Kdegkill\times{[kill]}
-</div>
-</html>
-After trxC promoter is turned on by ROSoxyR complex and the downstream genes – LacI, LuxI, and LuxR starts to produce, which blocks the CI promoter and turns on the LuxCI hybrid promoter. After that, the gene encoded with kill protein downstream LuxCI hybrid promoter also begins to transcribe.
+==Results==
+[[File:NYMU_defensin normal.png]]
-This equation shows production rate of mRNAkill. PoPSLuxcI represents the promoter strength of promoter trxC and LuxcI hybrid promoter, which is measured by the rate of RNApolymerase binding to the starting site of DNA transcription; <html><span lang="latex">\frac{1-[ LuxRAHL]^nLuxRAHL}{ KdLuxRAHL ^nLuxRAHL + [LuxRAHL]^nLuxRAHL} </span></html>represents the hill effect of activator LuxRAHL to LuxcI hybrid promoter; <html><span lang="latex">\ frac{1-[CI leak]^nCI}{ KdCI^nCI+[CI]^nCI}</span></html> represents the hill effect of repressor CI, while <html><span lang="latex"> frac{1-{1-[CI leak]^nCI}}{ KdCI^nCI+[CI]^nCI}</span></html> takes CI leaking into consideration (promoter pLac will not close thoroughly even though CI exists).
+Figure1: This picture shows kill protein concentration to time under pLux/CI hybrid promoter’s control and LuxR, LuxI’s positive feedback. The result indicates that kill protein concentration will reach its effective level and kill ''Nosema'' in time (36 hours after ''Nosema'' infection)
-For the section of the equation, <html><span lang="latex">\frac{1-[ LuxRAHL]^nLuxRAHL}{ KdLuxRAHL ^nLuxRAHL + [LuxRAHL]^nLuxRAHL}\times \frac {1-[CI leak]^nCI}{ KdCI^nCI+[CI]^nCI}\times {PoPSLuxcI} \times \frac{N}{V} </span></html>
+[[File:NYMU_defensin witout ROS.png]]
-represents the synthesizing rate of mRNAkill under the influence of LuxRAHLCI repression and leaking, and LuxcI hybrid promoter; <html><span lang="latex">-kdegmRNA\times [mRNALuxR]</span></html>   represents the degrading rate of mRNAkill.
+Figure2: This picture is a control group model. It shows that without ''Nosema'' infection (ROS concentration=0), ethanol production is low enough to be ignored.
-As for explanations of equation7, 8, 9, 15, 16, 17, 20, please see the ethanol wiki for more information.
+[[File:NYMU_defensin only CI.png]]
-==Results:==
+Figure3: This is the picture of kill protein concentration to time under promoter CI’s control only(control group)without pLux/CI's regulation.The result shows that kill protein's full expression is too low to kill ''Nosema''.
-[[File:NYMU_kill.jpg]]
-After Nosema invasion (t=0), concentration of kill protein will reach its constant level, which is <html><span lang="latex">6.5\times 10^(-9)M</span></html>. The effective concentration of kill protein to kill Nosema is <html><span lang="latex">6.7\times 10^(-10)M</span></html>, which is near the level we have modeled. Besides, the time to reach this concentration is 24 hours after Nosema invasion. Since three days after Nosema invasion, bees will start to spread Nosema, the constant level of kill protein (which is a little higher than the effective one)and the time it needs to reach this level (which is a little bit shorter than )shows that Bee. Coli can save the bees timely.
+==Discussion==
-[[File:NYMU_ethanol.jpg]]
+Comparing to the ROS-level control group where ROS remains low when there’s no ''Nosema'' infection, kill protein production is highly boasted upon sensing ''Nosema''.
-Similarly, the effective concentration of ethanol to kill bees is <html><span lang="latex">2.4\times 10^(-6)M</span></html> and the time to reach this concentration is 72 hours after Nosema invasion. Since kill protein will kick on two days after Nosema invasion, the open time of ethanol (four days after Nosema invasion) is what we desired. That is, because ethanol is the last end of preventing Nosema from spreading (it kills the infected bees as well), we want the open time of ethanol to be later than kill protein open time.
+The result indicates that kill protein concentration will reach its effective level and kill ''Nosema'' in time (36 hours after sensing ''Nosema'' infection, within the 5-day deadline when'' Nosema'' spores proliferates and the bee become incurable and infectious to other bees), at the given ROS changes and the enhanced OxyR expression [[(to learn more about our sensor model, click here)]].
-	The benefits of our circuit are that the dissociation constant of T7 (KdT7) is small, which means it is sensitive (will swiftly open right after T7 polymerase reaches threshold) and that we use positive feedback to attain the goal of open this device like a switch.
+==Circuit design and improvement==
+Upon using pCI to negatively regulate kill protein production, results show that kill protein's full expression while no CI present is too low to kill ''Nosema''. To solve this problem we subsidized CI promoter into pLux/CI promoter for additional positive feedback to boast kill protein’s production. As the modeling results regarding the modified circuit shown, the consequences are positive. [[(click here to learn more about our circuit)]]
 ==Parameters:==
 {|class="wikitable" !Model!!Parameter!!Description!!Value!!Unit!!Reference
@@ Line 660: / Line 540: @@
 | colspan="1" style="text-align: center;" | b
-| colspan="1" style="text-align: center;" | Infection rate constant of Nosema ceranae to the suspected
+| colspan="1" style="text-align: center;" | Infection rate constant of ''Nosema'' ceranae to the suspected
 | colspan="1" style="text-align: center;" | 24/75
 | colspan="1" style="text-align: center;" | Period(days)-1