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Position: Home > Articles > Cloning and Expression of Hai-Lan Chicken Interleukin-18 in E.coli and Preparation of Its Antiseru Journal of South China Agricultural University 2008,29 (1) 94-97

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

作  者:
赵丽;崔保安;陈红英;宋凌云;方忠意;郑兰兰
单  位:
河南省兽药监察所;河南农业大学牧医工程学院
关键词:
鸡白细胞介素18;成熟蛋白;克隆;原核表达
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.
译  名:
Cloning and Expression of Hai-Lan Chicken Interleukin-18 in E.coli and Preparation of Its Antiseru
作  者:
ZHAO Li1,2,CUI Bao-an1,CHEN Hong-ying1,SONG Ling-yun1,FANG Zhong-yi3,ZHENG Lan-lan1(1 College of Animal Husbandry and Veterinary,He'nan Agricultural University,Zhengzhou,450002,China; 2 Department of Pharmaceutical Engineering,College of Zhengzhou Animal Husbandry Engineering,Zhengzhou 450011,China;3 He'nan Institute of Veterinary Drug Control,Zhengzhou 450002,China)
关键词:
chicken IL-18;mature protein;clone;prokaryotic expression
摘  要:
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 nucleotide sequence of this ChIL-18 mature protein gene was 5l0 bp including the stop coden and,pQE30-ChIL18 was obtained by subcloning 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.

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