Cloning of mutA and expression of its coding protein
- VernacularTitle:变形链球菌CH43变链素Ⅰ基因片段的克隆及表达
- Author:
Xin SHI
;
Longxing NI
;
Yu TIAN
- Publication Type:Journal Article
- Keywords:
Streptococcus mutan;
Dental caries;
Mutacin;
mutA;
Cloning;
Expression
- From:
Journal of Practical Stomatology
2001;0(03):-
- CountryChina
- Language:Chinese
-
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 ion:mutA of Ms CH43 can be cloned and expressed in E.coli DH 5?.
