猪干扰素调节因子7的克隆及原核表达

doi:10.11843/j.issn.0366-6964.2014.08.002

畜牧兽医学报 ›› 2014, Vol. 45 ›› Issue (8): 1213-1217.doi: 10.11843/j.issn.0366-6964.2014.08.002

猪干扰素调节因子7的克隆及原核表达

张雪梅，杜丽丽，刘福涛，王江，郭豫杰，李宏基，杨国宇^*

(河南农业大学农业部动物生化与营养重点开放实验室，郑州 450002)

收稿日期:2014-02-24 出版日期:2014-08-23 发布日期:2014-08-23
通讯作者: 杨国宇，教授，博士，主要从事动物生物化学研究，E-mail：haubiochem@163.com
作者简介:张雪梅（1990-），女，河南济源人，硕士生，主要从事动物生物技术研究，E-mail：775347290@qq.com
基金资助:
农业部948重点计划（2011-G35）；河南省重点科技攻关（112102310705）

Cloning and Prokaryotic Expression of the Porcine Interferon Regulatory Factor 7 (IRF7)

ZHANG Xue-mei，DU Li-li，LIU Fu-tao，WANG Jiang，GUO Yu-jie，LI Hong-ji，YANG Guo-yu^*

(Key Laboratory of Animal Biochemistry and Nutrition of Ministry of Agriculture，Henan Agricultural University，Zhengzhou 450002，China)

Received:2014-02-24 Online:2014-08-23 Published:2014-08-23

摘要/Abstract

摘要：

为了进一步研究猪IRF7基因所编码蛋白质的生物学功能，本研究以猪十二指肠cDNA为模板扩增IRF7基因ORF区；将该基因克隆至原核表达载体pET-21b(+)中，获得pET-21b-IRF7重组质粒。经测序鉴定后，将重组质粒转化E.coli BL21 (DE3)，IPTG诱导表达His-IRF7融合蛋白，并利用Western blot进行鉴定。结果表明：（1）成功克隆了1 461 bp IRF7 ORF区；（2）His-IRF7融合蛋白在裂解菌液的上清和沉淀中均有表达，相对分子质量为60.5 ku。结果提示，运用大肠杆菌表达系统成功表达IRF7融合蛋白，为猪IRF7基因相关功能研究提供了基础材料。

Abstract:

To gain further insight into the biological functions of IRF7 gene，the open reading frame of the target gene was amplified from the cDNA of porcine duodenum，and cloned into the prokaryotic expression vector pET-21b(+) to generate a pET-21b-IRF7 recombinant plasmid.After verification by sequencing，the pET-21b-IRF7 was constructed，and then transformed into E.coli BL21(DE3).The His-IRF7 fusion protein was induced by IPTG and then identified by Western blot.The results shown that：(1) The open reading frame of 1 461 bp was successfully cloned.(2)His-IRF7 fusion protein was efficiently expressed in the lysate in the form of both soluble cysteine and inclusion bodies,with a molecular weight of about 60.5 ku.These results indicated that IRF7 fusion protein was successfully expressed in E.coli BL21(DE3)，which lays the foundation for the related function studies of porcine IRF7.

中图分类号:

S828
S813.3

张雪梅，杜丽丽，刘福涛，王江，郭豫杰，李宏基，杨国宇. 猪干扰素调节因子7的克隆及原核表达[J]. 畜牧兽医学报, 2014, 45(8): 1213-1217.

ZHANG Xue-mei，DU Li-li，LIU Fu-tao，WANG Jiang，GUO Yu-jie，LI Hong-ji，YANG Guo-yu. Cloning and Prokaryotic Expression of the Porcine Interferon Regulatory Factor 7 (IRF7)[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(8): 1213-1217.

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猪干扰素调节因子7的克隆及原核表达

Cloning and Prokaryotic Expression of the Porcine Interferon Regulatory Factor 7 (IRF7)

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