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| Cloning,Prokaryotic Expression and Function Analysis of GmALDH 3-1 from Danbo Black Soybean |
| ZHANG Le, CHEN Xuan-qin, WANG Lin, CHEN Li-mei, LI Kun-zhi |
| College of Life Science and Technology, Kunming University of Science and Technology, Kunming 650500, China |
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Abstract Aldehydes were one kind of toxic substance with high activity of chemical reactions. In the body of organism, aldehydes were chiefly generated by the lipid peroxidation, amino acid oxidation, and protein glycation. ALDHs (aldehyde dehydrogenase),with a variety of aldehydes as the substrate, encoded one kind of proteins responsible for the oxidation of aldehyde to carboxylic acid, can be found in plants, animals, and microorganism. The ALDH cDNA sequence of 'Danbo black soybean’ was amplified by RT-PCR. Prokaryotic expression vector pGEX-4T-1-ALDH3-1 was constructed and transformed into E.coli BL21 for expression. The effect of different concentrations of IPTG, time, and temperature on the ALDH protein expression was studied. The results showed that maximum recombinant protein was expressed in 6h at 28℃ with induction of IPTG, while the concentration of IPTG has little effect on the expression level of ALDH3-1. The transformed bacteria and control were cultured in liquid LB medium containing different concentration of Al, Cu, and Cd ions The growth curve of them was detected. The results show that ALDH3-1 has a certain tolerance to the mental ions mentioned above in vivo. These results provided foundation for the further investigation on ALDH3-1 structure and biological function.
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Received: 03 December 2012
Published: 25 May 2013
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