抗菌肽PekⅡ基因在大肠杆菌中的融合表达及初步纯化  

Fusion Expression of PekⅡ Gene in Escherichia coli and Its Primary Purification

作　　者：朱杰[1,2] 陈素华[2] 宋珊珊[2,3] 董先智[2] 

机构地区：[1]安徽农业大学生命科学学院,安徽合肥230036 [2]中国科学院生物物理研究所,北京100101 [3]中国海洋大学海洋生命学院,山东青岛266003

出　　处：《现代生物医学进展》2009年第4期676-678,738,共4页Progress in Modern Biomedicine

摘　　要：目的:研究抗菌肽PekⅡ基因在大肠杆菌中的融合表达并初步纯化。方法:根据大肠杆菌密码子的偏好性,人工设计并合成2段核苷酸序列,退火获得PekⅡ基因;将此基因克隆到原核表达载体pGEX-4T-2中,构建成抗菌肽基因PekⅡ融合表达载体pGEX-PekⅡ,转化至大肠杆菌BL21中,用IPTG进行诱导。取超声破碎后的上清经Glutathione SepharoseTM 4亲和层析得到纯化融合蛋白。结果:PCR和测序表明已获得正确PekⅡ编码基因,SDS-PAGE显示29kD处有特异性的蛋白条带出现,纯化得到的GST-PekⅡ经MALDI-TOF MS分析得出其相对分子质量为29553.03。结论:抗菌肽PekⅡ基因在大肠杆菌中的融合表达获得成功,初步纯化得到纯品。Objective： To investigate the fusion expression of Pek Ⅱ gene from antimicrobial peptides in Escherichia coli and its primary purification. Methods： Based on the preference condon of E. coli, two gene sequence were designed and synthesized. The gene ofPek Ⅱ was obtained by anneal. The fragment was inserted into pGEX-4T-2 and the expression plasmid pGEX-Pek Ⅱ was constructed. After sequencing, the gene was transferred into E. coli BL21 and induced by IPTG. Then the fusion proteins were purified by affinity chromatography with Glutathione SepharoseTM 4 after ultrasonication. Results： The result of PCR and sequencing showed that the correct gene was obtained, and the specific protein brand was at 29 kD by SDS-PAGE. The result of MALDI-TOF MS showed that the purified protein had a brand with 29553.03 Da. Conclusions： The Pek II gene was expressed in E.coli successfully, and the recombinant protein was greatly purified.

关 键 词：抗菌肽 大肠杆菌 融合蛋白 表达 

分 类 号：R78[医药卫生—口腔医学] R916[医药卫生—临床医学]