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.
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)
关键词
鸡γ-干扰素
原核表达
密码子偏嗜性
Chicken interferon gamma
Prokaryotic expression
Condon preference
