Cloning of mutA and expression of its coding protein / 实用口腔医学杂志

ABSTRACT

Objective:To obtain mutA gene of Strepto c occus mutans (Ms),and to express it in E.coli DH5?.Methods: mutA gene was amplified by PCR with specific primers from genome of Ms CH43 strain. After sequencing, the gene segment was inserted into vector pProEX and expressed in E.coli DH5?.The protein expression was induced by ITPG an d the protein products were examined by 180 ml/L SDSPAGE electrophorosis. Results:The length of PCR product was 147 bp and was identical to mu tA gene reported by GenBank.The mutA gene product was expressed in E.col i DH5? with Mr of 5.7?10 3.The maximum mutA protein product amount (20% of the total bacterial protein) was obtained when the A 600 value of DH5? was 1.666,IPTG concerntration 1.0 mmol/L and induction time 6 h.Conclus ionmutA of Ms CH43 can be cloned and expressed in E.coli DH 5?.