Cloning and expression of α-glucosidase gene from Thermotoga neapolitana

To produce a kind of Thermotoga neapolitana DSM 4359 α-glucosidase(TNAG) in large quantity which GenBank registration number was lcl27401,the gene was cloned and expressed in Escherichia coli.First,the genome DNA of T.neapolitana DSM 4359 as a template and through the PCR amplification target gene and complete the T cloning,the results of database contrast in NCBI indicated: the similarity of amino acid sequences of target gene and Thermotoqa sp.RQ2 was more than 99%.Then the target gene was connected to expression vector pET-32a(+) and E.coli Rosetta was imported with isopropyl β-D-1-thiogalactopyranoside(IPTG) induction expression.A obvious protein band of about 72 KDa was displayed by sodium dodecyl sulfate polyaclamide gel electrophoresis(SDS-PAGE).After heat treaments,the enzyme's optimal temperature and pH were 80℃ and about 5.0.