兔IFN-γ基因的克隆、遗传进化分析及其表达  

Cloning,phylogenetic evolution and expression of rabbit IFN-γ gene

作　　者：孟庆玲[1] 乔军[1] 才学鹏[1] 闫鸿斌[1] 骆学农[1] 景志忠[1] 

机构地区：[1]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室

出　　处：《甘肃农业大学学报》2008年第4期7-11,共5页Journal of Gansu Agricultural University

基　　金：中国农业科学院兰州兽医研究所所长基金项目( 2006-02);甘肃省农业生物技术研究与应用开发项目(5CS055-A43-057)

摘　　要：运用RT-PCR技术对用ConA刺激后的中国白兔外周血淋巴细胞（PBMCr）进行扩增。将纯化后的PCR产物克隆人pMD18-T中进行序列测定。并进行遗传进化分析．结果发现：该基因全长501bp，编码167个氨基酸．其中前20个氨基酸残基构成信号肽序列．与不同物种IFN-γ基因相比，核苷酸和推导的氨基酸序列有一定的差异．在推导的中国白兔IFN-γ氨基酸序列中。在41～43、108～110和117～119位存在3个潜在的N-联糖基化位点，同时存在3个Cys残基．转化重组质粒pETIFN-γ的大肠杆菌BL21（DE3）经IPTG诱导后，重组菌菌体裂解物经SDS-PAGE电泳可检测到相对分子量为22．6kDa的重组蛋白．经凝胶薄层扫描．重组蛋白表达量可占菌体蛋白的19．2％．Chinese white rabbit IFN-γ gene was amplified by RT-PCR method from the PBMCr stimulated by ConA. Then the PCR product was purified and ligatured with pMD18-T vector. The positive recombinant was used for sequencing. The complete length of white rabbit IFN-γ gene was 501 bp ,which encoded 167 amino acids. The front 20 amino acids consist of signal peptide. Compared with the IFN-γ genes of other species,there were some differences in the nucleotide sequence and deduced amino acid sequence. There were 3 N-glycosylation sites and 3 Cys in the deduced amino acid sequence. The host E. coli strain BL21 （DE3） transformed with recombinant pETIFN-γ can express a recombinant protein with molecular weight of 22.6 kDa under the induction of IPTG,which amounted to 19.2% in the total protein of the induced bacteria by the assaying of gel scanning.

关 键 词：中国白兔 IFN-Γ基因 克隆 序列分析 表达 

分 类 号：S852.4[农业科学—基础兽医学] Q785[农业科学—兽医学]