根癌农杆菌attM基因的克隆及表达  

Cloning and Expression of attM Gene from Agrobactrium tumefacien

作　　者：马宏[1] 贾振华[1] 张岩峰[1] 张星炜[2] 房宝玲[3] 宋水山[1] 

机构地区：[1]河北省科学院生物研究所,河北石家庄050081 [2]邯郸市环境保护研究所,河北邯郸056002 [3]河北省邯郸学院,河北邯郸056005

出　　处：《生物技术》2013年第1期8-11,共4页Biotechnology

基　　金：河北省科学院青年创新基金项目(No.11330);河北省科学技术研究与发展计划项目(No.11215504D)资助~~

摘　　要：目的:利用γ-丁内酯在基因工程菌中合成聚-4-羟基丁酸酯。方法:利用PCR技术克隆内酯水解酶attM基因,进行体外表达,并导入质粒pKSSE5.3,构建新的工程菌,进行摇瓶培养。结果:测序结果显示DNA序列全长1 077 bp,包含771bp的开放阅读框,编码256个氨基酸。携带attM基因的表达载体能够进行正常表达,内酯酶活性为5.82U/mL;在摇瓶培养条件下,工程菌E.coli XL1-Blue(pKSSE5.3M9)的培养液中P(4HB)占细胞干重的含量为29%。结论:成功克隆了attM基因,新构建的工程菌能利用γ-丁内酯为碳源合成聚-4-羟基丁酸酯。Objective： Production of Poly （4 - hydroxybutyric acid） from y - butyrolactone by recombinant Escherichia coli. Method： The attM gene from Agrobactrium tumefacien was cloned by PCR, expressed in vitro and assembled onto the plasmid pKSSE5. 3 which was used to synthesize the homopolyester P（4HB）. Result：The result of sequencing showed that the full length of attM was 1 077bp which contai- ning an opening reading frame of 771 bp and encoding 256 amino acid. The plasmid integrated attM gene could expressed properly. The en- zyme activity of AttM was 5. 82U/mL. P（4HB） accumulation was 29% of dry cell weight,when cells were cultivated in mineral salts M9 medium plus y- butyrolactone as carbon source. Conclusion：The attM gene was cloned. P （4HB） accumulation was compounded when the new recombinant was cultivated with ~ -butvrolactone as carbon source.

关 键 词：根癌农杆菌 attM基因 Γ-丁内酯 4-羟基丁酸 聚-4-羟基丁酸酯 重组大肠杆菌 

分 类 号：Q786[生物学—分子生物学]