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| <section><h3>Cloning of Module 1 in pZS21MCS:</h3> | | <section><h3>Cloning of Module 1 in pZS21MCS:</h3> |
− | <h4>For cloning of construct 1, Ptet- RBS1- T7 RNA polymerase –terminator 1-terminator 2 and construct 2, P<sub>T7</sub>-RBS2- Chromoprotein II- RBS3-tetR-terminator3. Different parts like Ptet, P<sub>T7</sub>, RBS, T7 RNA polymerase, Chromoprotein II, tetR and terminator parts are taken from BioBricks library and are amplified by PCR. Different amplified PCR products are fused together through splice overlap extension PCR.</h4> | + | <h4>Cloning of construct 1 --> Ptet - RBS1 - T7 RNA polymerase - terminator 1 - terminator 2</h4> |
| + | <h4>Cloning of construct 2 --> P<sub>T7</sub> - RBS2 - Chromoprotein II - RBS3 - tetR - terminator3.</h4> |
| + | <h4>Different parts like Ptet, P<sub>T7</sub>, RBS, T7 RNA polymerase, Chromoprotein II, tetR and terminator parts are taken from BioBricks library and are amplified by PCR. Different amplified PCR products are fused together through splice overlap extension PCR.</h4> |
| <h3>Cloning of Module 2 in pACYC177 :</h3> | | <h3>Cloning of Module 2 in pACYC177 :</h3> |
| <h4>For cloning of construct 1 of module 2, Pmar-RBS4- ToxR-terminator 4 and construct 2, Pctx-RBS5- chromoprotein I- RBS6-TetR- terminator5. Different parts like ToxR and chromoprotein I are PCR amplified from BioBricks library. Other parts like- Pmar is amplified from <i>E. coli</i> genome (BW25113), and terminator 4 is cloned from plasmid (modified pAH125). Parts are amplified by PCR and different parts are fused together by splice overlap extension PCR.</h4> | | <h4>For cloning of construct 1 of module 2, Pmar-RBS4- ToxR-terminator 4 and construct 2, Pctx-RBS5- chromoprotein I- RBS6-TetR- terminator5. Different parts like ToxR and chromoprotein I are PCR amplified from BioBricks library. Other parts like- Pmar is amplified from <i>E. coli</i> genome (BW25113), and terminator 4 is cloned from plasmid (modified pAH125). Parts are amplified by PCR and different parts are fused together by splice overlap extension PCR.</h4> |