鸡γ-干扰素的克隆·表达和序列分析.pdfVIP

安徽农业科学，Journal ofAnhui Agri．Sci．2008，36(17)：7138—7140 责任编辑 陈玉敏 责任校对 卢瑶 鸡 -干扰素的克隆·表达和序列分析 谢昆 ，王传铭t，林丽飞1，王栋t，李河1 (1．红河学院生命科学与技术学院，云南蒙自661100；2．云南省天然药物与化学生物学重点实验室，云南蒙自 661100) 摘要 [目的]为鸡 一干扰素的进一步研究奠定基础。[方法]根据GenBank发表的鸡 一干扰素核苷酸序列，设计l对特异性引物。利 用RT-PCR技术从gonA诱导培养的鸡脾脏淋巴细胞中克隆鸡 ．干扰素基因，并在IPTG诱导下在大肠杆菌中表达重组茵。[结果]克 隆的鸡 ．干扰素基因全长495 bp，编码165个氨基酸，与国外克隆的鸡 一干扰素基因序列的同源性为100％。鸡 一干扰素蛋白合有 l2个ot螺旋，3个B折叠，7个B转角。它具有强抗原性，而第16～23位氨基酸无抗原性。重组蛋白的分子量约18 kD，其经IPTG诱导 表达4 h后，表达量最高，表达产率高达25％。[结论]鸡 ～干扰素具有一定的抗原性，预示其在免疫反应中起一定作用。 关键词 鸡 ．干扰素；克隆；表达 中图分类号 $831 文献标识码 A 文章编号 0517—6611(2oo8)17—07138—03 Cloning，Expression and Sequence Analysis of Chicken Interferon-v X皿Kun etal(College ofLife Science andTechnology，HongheUniversity，Mengzi．Yunnan 661lOO) Abstract 『Objective1 rhe aim of the research WaS to lay the foundation for further study on chicken interferon ．『Method1 According to the nucleotide sequence of chicken interferon-~／that was published on GenBank wehsite a pair of specific primers were designed．Chicken interferon一 was cloned from the lymphocyte in chicken spleen through ConA induced culture by using RT-PCR technology．And the recombinant bacteria were expressed in Escherichia col through lPTG induction．fResultl The cloned chicken interferon~with total length of 495 bp coded 165 amiI10 acids and its homology with the foreign cloned chicken interferon gene sequence was 100％．Chicken intefferon protein contained twelve d helices，three B sheets and seven B turs．It had a strong antigenieity，but it had no antigenieity on the 16 一23 amino acid．The moleeular wright of the recombinant protein was about 18 kD and the expression 1evel was highest after IPTG induction expression for 4 h．with the expression yield reaehing 25％．『Conclusion1 Chicken interferon一 had certain antigenicity and this indicted that it had certain fuction in the immune response． KeY words Chicken interferon-~／；Cloning；Expression 近年来，随着病原菌新毒株、变异株的不断出现，我