油菜偏嗜性ChIFN-γ基因合成及原核表达  

Gene Synthesis and Prokaryotic Expression of ChIFN-γ with the Brassica napus codon usage optimization

作　　者：许文钊[1] 吴拥军[1,2] 赵德刚[1,2] 田斌[2] 

机构地区：[1]贵州大学农业生物工程省级重点实验室,贵阳550025 [2]贵州大学生命科学学院,贵阳550025

出　　处：《山地农业生物学报》2009年第3期250-254,共5页Journal of Mountain Agriculture and Biology

基　　金：国家科技支撑计划项目资助(2007BAD59B06);科技部国际科技合作项目资助(2007DFA31260)

摘　　要：依据鸡γ-干扰素基因(ChIFN-γ)的氨基酸序列,采用油菜偏嗜性的密码子,人工合成ChIFN-γ,构建原核表达载体pET32a-ChIFN-γ,并转化到E.coliBL21中。1 mmol.L-1IPTG 37℃诱导表达12h ChIFN-γ表达量达到菌体总蛋白的37%。SDS-PAGE电泳检测表明,ChIFN-γ重组蛋白分子量约为33kDa。研究为下一步制备ChIFN-γ抗体进行真核表达研究奠定了基础。According to the amino acids sequence of chicken interferon gamma （ChIFN-γ） gene and the codon usage bias of Brassica napus, the gene encoded ChIFN-γ was successfully synthesized. The gene was cloned into vetor pGM - T. The sequencing result suggested that the cloned sequence coincided with the designed sequence. Subsquently, the gene was cloned into vector pET32a and into Escherichia coli BL21.ChIFN-γ was expressed at yield about 37% of total protein in Escherichia coli BL21strain by 1.0 mmol .L-1 IPTG induced for 12h at 37℃. A specific expression band with a relative molecular weight 33 kDa was observed by SDS - PAGE. This research had established a platform for the preparation of antibody and eukaryotie expression of ChIFN-γ.

关 键 词：鸡γ-干扰素 原核表达 密码子偏嗜性 

分 类 号：Q786[生物学—分子生物学]