期刊论文

Construction of the Expression System for Gpd Gene From Treponema Pallidu and Antigenicity Identification of the Recombinant Protein

中文

中南医学科学杂志

2095-1116

2005

33

1

29-32,36

10.3969/j.issn.2095-1116.2005.01.008

湖南省教育厅重点资助项目 （ 0 0 2A0 46）； 湖南省卫生厅科研基金 （B2 0 0 3 -0 85 ）；

本校为第一机构

目的　构建梅毒螺旋体 （Treponemapallidum ,Tp）外膜蛋白Gpd基因的原核表达载体 ,检测其表达产物的免疫原性 ,并比较梅毒螺旋体各菌株Gpd基因序列的同源度。方法　从TpNichols株基因组模板中PCR扩增Gpd基因 ,与GenBank登录的序列做blast比较 ,定向克隆构建原核表达重组体pET2 8b（ + ） -Gpd ,转入大肠杆菌ril表达菌 ,SDS -PAGE分析重组蛋白的表达 ,Western -blot检测重组蛋白的免疫原性。结果　载体上所连目的基因片段序列与GenBank登录的Nichols株Gpd基因序列完全一致 ,同其他病原性密螺旋体菌株登陆序列比较同源度为 98%～ 1 0 0 %。SDS -PAGE检测诱导产物显示有一Mr约为 41kDa的特异蛋白带...

Objective To construct the recombinant plasmid of Procaryotic expression containing gpd gene from Treponema Pallidum(T.pallidum) and identify immunity of the expression product. Methods gpd gene was amplified from the genomic DNA of T.pallidum by polymerase chain reaction (PCR),and the gene was subcloned into appropriate site of pET28b(+) vector.After identification by sequencing and restrictive enzymes digestion,the recombinant plasmid was transfected into E.col ril strain.The expressed protein was identified by western-blot. Results The targe...