摘要
根据Genebank中已发表的INF-γ蛋白cDNA序列保守区设计一对特异性引物,应用RT-PCR技术从Con A活化的牛外周血单核细胞中扩增到编码牛INF-γ蛋白基因,其大小为510 bp左右。将该基因克隆到pMD18-T载体中,测序结果与Genebank上发表的INF-γ基因序列进行比较,其同源性为99.8%。将该基因从重组pMD18T-IFN质粒中克隆到表达载体pcDNA3.1(+)质粒中,构建真核表达重组质粒,经酶切和PCR鉴定后表明构建的重组质粒为阳性。
According to the encoding sequence of the INF-γprotein in Genebank and the multiple cloning sites characteristic of the eukaryotic expression vector peDNA3.1 （＋）, appropriate primers specific to INF-γgene were designed. The target DNA fragments were obtained by RT-PCR amplification and then were cloned into the pMD18-T vector. After identification by restriction enzyme digestion and sequencing analysis, the specific DNA fragments were cloned into the eukaryotic expression vector pcDNA3.1 （＋）. The positive recombinant plasmids were named pcDNA3.1 （＋）-INF after identification by restriction enzyme digestion and PCR amplification. Results showed that recombinant expression plasmids pcDNA3.1（＋）-INF were successfully constructed.
出处
《黑龙江八一农垦大学学报》
2010年第1期53-57,共5页
journal of heilongjiang bayi agricultural university
基金
校硕士启动基金资助项目(S2005-34)
黑龙江省教育厅资助项目(11531262)
关键词
干扰素-Γ
真核表达
载体构建
INF-γ
eukaryotic expression
vector construction
