死亡肽基因的合成及在大肠杆菌中的表达  

Cloning and fusion expression of the antimicrobial peptide thanatin gene in Escherichia coli

作　　者：翁宏飚[1] 牛宝龙[2] 孟智启[2] 徐孟奎[1] 

机构地区：[1]浙江大学动物科学学院,杭州310029 [2]浙江省农业科学院蚕桑研究所,杭州310021

出　　处：《昆虫学报》2003年第1期114-117,共4页Acta Entomologica Sinica

基　　金：浙江省重点项目 ( 0 2 110 2 0 86 )

摘　　要：将人工合成的寡核苷酸片段 ,通过PCR扩增得到死亡肽 (thanatin)基因 ,并将其克隆到表达载体pGEX 3X中 ,序列分析结果正确。经IPTG诱导 ,在大肠杆菌BL2 1中进行高效可溶性表达 ,表达量可达 2 0 %以上。融合蛋白通过GST亲和层析纯化 ,用肠激酶酶解表达产物 ,用SephadexG 2 5初步纯化得到具有抗菌活性的死亡肽。The thanatin gene was obtained and inserted into expression vector pGEX-3X by a DNA recombinant method which was checked by nucleotide sequencing. The fusion protein of GST-thanatin was produced by IPTG induction in Escherichia coli (BL21). The expression level was about 20%. The fusion protein was purified by GST affinity chromatography and digested by enterokinase. Partly purified with Sephadex G-25, the final product, thanatin exhibited antimicrobial activity.

关 键 词：死亡肽 融合表达 大肠杆菌 

分 类 号：Q516[生物学—生物化学] R378.21[医药卫生—病原生物学]