摘要
参照IBRV基因组序列,设计合成1对特异性引物,扩增了长约846bp的gD基因片段。将目的片段定向克隆到pET-30a原核表达载体,酶切及测序鉴定均正确后,转化BL21(DE3)表达菌,经IPTG诱导得到了以可溶性形式表达的重组蛋白。重组蛋白纯化后,经免疫印迹检测证明具有良好的抗原性和特异性。本研究利用原核表达系统成功表达具有良好抗原性的gD蛋白,为IBRV的深入研究及诊断试剂的研制奠定了物质基础。
According to the published IBRV genome sequence,we designed and synthetized one pair of specific primers,amplied about 846 bp gD gene fragment.The fragment was cloned into prokaryotic expression vector pET-30a,after enzyme digestion and DNA sequencing were correct,the recomninated vector was transformed into BL21（DE3） bacteria and induced by IPTG,recombinant protein was expressed in partially soluble form.After purification of recombinant protein,Western blotting proved that it had good antigenicity and specificity.The present study using prokaryotic expression system had good antigenicity of gD protein,which laid a material foundation for IBRV further research and development of diagnostic reagents.
出处
《中国畜牧兽医》
CAS
北大核心
2013年第3期77-80,共4页
China Animal Husbandry & Veterinary Medicine
关键词
牛传染性鼻气管炎病毒
gD蛋白
原核表达
鉴定
infectious boviner rihinotracheitis virus
gD protein
prokaryotic expression
identification
