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| | <p>Plasmid Amplification for further construction; | | <p>Plasmid Amplification for further construction; |
| | <br>chose pRSFDuet-1, pACYCDuet-1 and pCDFDuet-1 for expression vector and pBluescript II KS（+） for connector.<p> | | <br>chose pRSFDuet-1, pACYCDuet-1 and pCDFDuet-1 for expression vector and pBluescript II KS（+） for connector.<p> |
| - | <h2 id ="JulyWeek2" >July Week2</h2> | + | <h2 id ="JulyWeek2" >July Week2</h2><br> |
| | <p>Construct the gene of our fusion protein: ssDsbA-FP-HL-Lgt-FL-TAL using splicing by overlap extension. | | <p>Construct the gene of our fusion protein: ssDsbA-FP-HL-Lgt-FL-TAL using splicing by overlap extension. |
| | We connected ssDsbA-FP-HL-lgt for one part of this fusion protein and FL-TAL for another. Then connect them together.<p> | | We connected ssDsbA-FP-HL-lgt for one part of this fusion protein and FL-TAL for another. Then connect them together.<p> |
| - | <h2 id ="JulyWeek3" >July Week3</h2> | + | <h2 id ="JulyWeek3" >July Week3</h2><br> |
| | <p> Connected ssDsbA-FP-HL-lgt by splicing by overlap extension. | | <p> Connected ssDsbA-FP-HL-lgt by splicing by overlap extension. |
| | Transform, colony picking plasmid extraction and digestion identification; Sequencing results showed accurate construction.<p> | | Transform, colony picking plasmid extraction and digestion identification; Sequencing results showed accurate construction.<p> |
| - | <h2 id ="JulyWeek4" >July Week4</h2> | + | <h2 id ="JulyWeek4" >July Week4</h2><br> |
| | <p> Use golden gate to connected TAL of 2012 Freiburg igem. | | <p> Use golden gate to connected TAL of 2012 Freiburg igem. |
| | Transform, colony picking plasmid extraction and digestion identification.<p> | | Transform, colony picking plasmid extraction and digestion identification.<p> |
| - | <h2 id ="AugustWeek1-2" >August Week1-2</h2> | + | <h2 id ="AugustWeek1-2" >August Week1-2</h2><br> |
| | <p>Test the conditions for the PCR. | | <p>Test the conditions for the PCR. |
| | Connected TAL, transform, colony picking plasmid extraction and digestion identification. Find with our electrophoresis band. Expression vectors and connector plasmid are confirmed by sequencing.<p> | | Connected TAL, transform, colony picking plasmid extraction and digestion identification. Find with our electrophoresis band. Expression vectors and connector plasmid are confirmed by sequencing.<p> |
| - | <h2 id ="AugustWeek3" >August Week3</h2> | + | <h2 id ="AugustWeek3" >August Week3</h2><br> |
| | <p>There are some problems about Freiburg’s parts. We can’t connected TAL in the right order. | | <p>There are some problems about Freiburg’s parts. We can’t connected TAL in the right order. |
| | So we design some new primes for PCR that can produce the right sequence.<p> | | So we design some new primes for PCR that can produce the right sequence.<p> |
| - | <h2 id ="AugustWeek4" >August Week4</h2> | + | <h2 id ="AugustWeek4" >August Week4</h2><br> |
| | <p>Design a few new ports for the fusion protein. | | <p>Design a few new ports for the fusion protein. |
| | Sequencing results showed accurate construction. Observe the FP using LSCM to confirm the fusion protein can locate on the membrane.<p> | | Sequencing results showed accurate construction. Observe the FP using LSCM to confirm the fusion protein can locate on the membrane.<p> |
| - | <h2 id ="SeptemberWeek1" >September Week1</h2> | + | <h2 id ="SeptemberWeek1" >September Week1</h2><br> |
| | <p>Try co-transformation: Prsf pacyc pBluescript . | | <p>Try co-transformation: Prsf pacyc pBluescript . |
| | Find the conditions of protein expression. | | Find the conditions of protein expression. |
| | Find the way to construct the TAL.<p> | | Find the way to construct the TAL.<p> |
| - | <h2 id ="SeptemberWeek2" >September Week2</h2> | + | <h2 id ="SeptemberWeek2" >September Week2</h2><br> |
| | <p>Find the enzymes for the application. | | <p>Find the enzymes for the application. |
| | Find the way to detect the substrate in these pathways. | | Find the way to detect the substrate in these pathways. |
| | Connector plasmid modification.<p> | | Connector plasmid modification.<p> |
| - | <h2 id ="SeptemberWeek3" >September Week3</h2> | + | <h2 id ="SeptemberWeek3" >September Week3</h2><br> |
| | <p>TAL gene synthesis. Construct the part with our new ports.<p> | | <p>TAL gene synthesis. Construct the part with our new ports.<p> |
| - | <h2 id ="SeptemberWeek4" >September Week4</h2> | + | <h2 id ="SeptemberWeek4" >September Week4</h2><br> |
| | <p> TAL gene synthesis.<p> | | <p> TAL gene synthesis.<p> |
| | </article> | | </article> |
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