Part:BBa_K2533033

RBS-gapA

It encodes glyceraldehyde-3-phosphate dehydrogenase.

Usage and biology

It encodes glyceraldehyde-3-phosphate dehydrogenase which could transform 3- phosphoglyceraldehyde into 1,3- diphosphoglycerate. With the overexpression of gapA, Shewanella could produce NADH more efficiently, which brings more electricity being produced.

Characterization

This is one section for NADH production part.

Figure1:RBS-gapA

DNA Gel Electrophoretic

To make sure that we get the target gene, we did the DNA gel electrophoretic to separate different gene. And here is the result.

Figure2:Verification of successful transformation of pYYDT-gapA

Our target genes are 1126bp, and as the marker is DS5000, we could be sure that the bright bands in this picture are our target genes.

Real-Time Quantitative PCR

To demonstrate that gapA could be overexpressed by engineered Shewanella, we did Real-Time Quantitative PCR.

File:T--HUST–China--2018-result-fig3.jpeg

Figure3:Relative expression level of gapA in engineered Shewanella Oneidensis MR-1.

There was no signal in bacteria which contained pYYDT so we chose pYYDT-gapA as standard quantity.