摘要
本研究旨在通过克隆鸡Mx全基因序列进而进行该基因的原核表达,获得具有生物学活性的蛋白。利用poly I:C诱导鸡胚成纤维细胞Mx基因表达,克隆了Mx基因全长cDNA序列,将开放阅读框(ORF)连接构建于表达质粒pGEX-4t-2中获得重组表达载体pGEX-Mx,转化Rosetta(DE3)菌株,经IPTG诱导后检测。表达产物检测显示该蛋白的相对分子量为75 ku。说明获得了Mx基因的高效表达,为进一步进行Mx基因的活性检测以及利用Mx蛋白进行抗病毒转基因的研究奠定了基础。
This experiment was conducted to study Mx gene expression in E. coli under the condition of cloning Mx gene complete sequence. Chicken embryo fibroblast (CEF) were treated with poly I:C to stimulate Mx gene expression. Full length cDNA of Mx was cloned. Recombinant expression vector pGEX-Mx including Mx ORF was constructed using pGEX-4t-2 expression system to transfer Escherichia coli Rosetta (DE3) strain. The result showed that the relative molecular weight of expressed protein was about 75 ku after induction with IPTG, suggesting that Mx gene has been efficiently expressed. The research laid a good foundation for further studying on bioactivity assay, exploring new way of antiviral medication and transgenic study.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2009年第7期978-981,共4页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
北京市自然科学基金资助项目(5051004)
国家高技术研究发展计划(863计划)项目(2007AA10Z183)
关键词
鸡
MX蛋白
克隆
表达
chicken
Mx protein
cloning
expression