海兰鸡白细胞介素18成熟蛋白基因的克隆及在大肠杆菌中的表达  

Cloning and Expression of Hai-Lan Chicken Interleukin-18 in E.coli and Preparation of Its Antiseru

作　　者：赵丽[1] 崔保安[1] 陈红英[1] 宋凌云[1] 方忠意[2] 郑兰兰[1] 

机构地区：[1]河南农业大学牧医工程学院 [2]河南省兽药监察所,河南郑州450002

出　　处：《华南农业大学学报》2008年第1期88-91,共4页Journal of South China Agricultural University

基　　金：“十五”国家食品安全重大攻关专项(2001BA804A30-11);河南省重大科技攻关项目(0223013800)

摘　　要：通过RT-PCR方法从用植物血凝素(PHA)活化60日龄海兰鸡的脾脏淋巴细胞中扩增出海兰鸡白细胞介素18(IL-18)成熟蛋白基因的cDNA,并将其克隆到pGEM-T Easy Vector载体上,DNA序列测定表明,克隆得到的海兰鸡ChIL-18 cDNA基因全序列包括终止密码子在内其编码区大小为510 bp,编码169个氨基酸多肽.将该基因片段克隆到原核表达载体pQE30构建重组质粒pQE30-ChIL-18,转化大肠杆菌M15,并用IPTG诱导.重组菌菌体裂解物SDS-PAGE可检测到相对分子质量为19 000的重组蛋白.Chicken interleukin-18 （ChIL-18） mature protein gene was amplified from PHA stimulated Hai-Lan chicken spleen cell by RT-PCR. PCR product was cloned into the pGEM-T vector. Sequencing analysis showed that the nueleotide sequence of this ChIL-18 mature protein gene was 510 bp including the stop eoden and,pQE30-ChIL18 was obtained by subeloning the encoding region of the ChIL-18 mature peptide gene into pQE30. The recombinant ChIL-18 （ rChIL-18 ） was expressed efficiently in pQE30-ChIL18-transformed M15 induced by IPTG. SDS-PAGE analysis showed that the recombinant mature protein had a relative molecular mass of approximately 19 000.

关 键 词：鸡白细胞介素18 成熟蛋白 克隆 原核表达 

分 类 号：S858.31[农业科学—临床兽医学]