Difference between revisions of "Part:BBa K1020012"

Revision as of 01:51, 29 October 2013

Alkane Selector for intracellular alkane production

We put the resistance gene encoding tetA under the regulation of PalkM. When PalkM-RNAP complex isomerizes due to the interaction between alkanes and alkR, downstream TetA gene will express. Then, TetA will transport tetracycline out of the cell so that the intracellular concentration of tetracycline is relatively stable. and the growth of cells will not be inhibited. Thus, we can turn alkane molecules into the output of the growth rate of cells.

We add alkanes outside the cell to test the function of TetA:

Characterization: Construct:Strains of No.60 are cultured in LB medium for about 12 hours. Add 12μg/mL Cm and 12μg/mL Tc respectively. After that, inoculate into 3 ml LB medium for an overnight cultures at 37 ℃ with 220rpm shaking.

Construct: Left: No.60: TetA downstream of Palkm and AlkR downstream of J23103, are cloned into vector PSB1C3 and there’s a alkane producing vector PSB3K3. Right: No.61: We construct a blank vector PSB3K3 without NPDC and AAR as blank control, and the other vector PSB1C3 is the same as No.60.

Result: Alkane producing E.coli can grow in tube but blank vector E.coli cannot grow.

Conclusion: Synthesized alkanes induce the expression of alkR, which enables E.coli to survive under the pressure of Tetracycline.

Sequence and Features