S-腺苷甲硫氨酸合成酶在大肠杆菌中的克隆与表达

Cloning and Expression of S-adenosyl-L-methionine Synthetase Gene in E.coli

目的:克隆与表达大肠杆菌S-腺苷甲硫氨酸合成酶的基因。方法:应用PCR技术从E.coliBL21(DE3)的总DNA中扩增出1.1kb的S-腺苷甲硫氨酸合成酶基因,将其连接到PGEMT载体上,测序证明正确后再将目的基因插入大肠杆菌高表达载体PET28a中,含有目的基因的重组质粒在大肠杆菌中进行表达与分析。结果:SAM合成酶在E.coliBL21(DE3)中的表达量达到23.6%,酶活达到4.17μmol/h.ml。结论:大肠杆菌表达出了具有生物活性的S-腺苷甲硫氨酸合成酶。

Objectives： To clone and express the S- adenosyl- L- methionine synthetase gene. Methods： The S- adenosyl- L- methionine synthetase gene was amplified with PCR from the genomic DNA of E. coli, and it was linked into PGEMT vector. After it was confiermed by sequencing analysis, it was inserted into the expression vector PET28α of E. coli. Then the recombined plasmid with target gene was expressed and analyzed. Results： The average expression of S- adenosyl - L- methionine synthetase in E. coli was up to 23.6%. And it＇s activity was up to 4. 17 μmol/l/ml. Conclusion：Tile S - adenosyl- L- rnethionine synthetase with biological activity was expressed in E. coli.