Cloning And Expression Of Human Aldehyde Dehydrogenase 2 Gene

Alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) play an important role in the in vivo alcohol metabolism together. The influence to central nervous system can be reduced, with rapid decomposition and enough metabolism of the alcohol ingested in the body full of the two enzymes. In Most of cases, ALDH is scare, while in vivo ADH is always expressed enough and evenly. Alcohol can't be completely decomposed to water and Carbon Dioxide, and redundant aldehyde will remain for the scarcity of ALDH, which leads to the palsy of central nervous system, drunken symptom, aldhl is a well-known high-level genetic polymorphic gene, and has a close affinity to the drinking behavior of the Asian. While the drug of de-alcoholic effects and preventing liver injury is demanded, it has a fine foreground of development and application.In this paper, the clone and expression of aldh2 gene from human was reported. aldhl gene was obtained from the genome DNA of human through PCR techniques, and cloned into pUCm-T vector, and determined by DNA sequencing system, aldhl gene fragment with restriction enzyme digesting site, which was generated by PCR with the plasmid pUCm-aldh2 as a template, inserted into an IPTG-inducible expression plasmid pET22b(+). The sequence of aldhl gene in the expression vector was verified by DNA sequencing analysis. The expression of ALDH-2 was achieved in E. coli with an IPTG induction. It makes a perfect expression in 37℃ and induced by lmmol/L IPTG whthin six hours. The target protein shared 30% of the total proteins of induced cells. The washed inclusion bodies were solublized with 8mol/L urea and purified using nickel affinity chromatography. The purified ALDH-2 was refolded by dilutedness in refolding solution with L-arginine. Then the activity of refolded ALDH-2 was determined. The results showed that protein has an ALDH specific activity of 331.7 U/mg.