| 猪链球菌2型次黄嘌呤核苷酸脱氢酶(IMPDH)重组蛋白的酶活性和免疫原性检测 |
| |
| 引用本文： | 张雪寒,何孔旺,周俊明,倪艳秀,俞正玉,吕立新.猪链球菌2型次黄嘌呤核苷酸脱氢酶(IMPDH)重组蛋白的酶活性和免疫原性检测[J].农业生物技术学报,2009,17(6). |
| |
| 作者姓名： | 张雪寒 何孔旺 周俊明 倪艳秀 俞正玉 吕立新 |
| |
| 作者单位： | 江苏省农业科学院兽医研究所,农业部动物疫病诊断与免疫重点开放实验室,国家兽用生物制品工程技术研究中心,南京,210014 |
| |
| 基金项目： | 国家重大基础研究发展规划(973)项目，江苏省自然科学基金项目 |
| |
| 摘 要： | 通过PCR方法从猪链球菌2型(Streptococcus suis)05ZYH33分离株基因组中扩增出次黄嘌呤核苷酸脱氢酶基因(inosine 5-monophosphate dehydrogenase,impdh),长度1 064 bp.PCR产物和pET28a载体分别经过EcoR Ⅰ和Xho Ⅰ双酶切,连接,成功地构建了重组表达质粒pET28a-impah,并转入大肠杆菌(Escherichia coli) BL21(DE3)中,经过1 mmol/L IPTG诱导获得表达,蛋白大小35kD.表达蛋白具有良好的抗原性和酶活性.蛋白经过亲和层析纯化后,在37℃,pH7.0～9.0表现出较强酶活性,NADH A_(340)介于0.662～0.816之间.
|
| 关 键 词： | 猪链球菌 次黄嘌呤核苷酸脱氢酶(IMPDH) 克隆 酶活性 |
Detection of Antigenicity and Enzymatic Activity of Recombinant Inosine5-monophosphate Dehydrogenase(IMPDH)of Streptococcus suis Serotype 2 |
| |
| ZHANG Xue-han,HE Kong-wang,ZHOU Jun-ming,NI Yan-xiu,YU Zheng-yu,LV Li-xin.Detection of Antigenicity and Enzymatic Activity of Recombinant Inosine5-monophosphate Dehydrogenase(IMPDH)of Streptococcus suis Serotype 2[J].Journal of Agricultural Biotechnology,2009,17(6). |
| |
| Authors: | ZHANG Xue-han HE Kong-wang ZHOU Jun-ming NI Yan-xiu YU Zheng-yu LV Li-xin |
| |
| Institution: | ZHANG Xue-han,HE Kong-wang,ZHOU Jun-ming,NI Yan-xiu,YU Zheng-yu,LV Li-xin (Institute of Veterinary Medicine,Jiangsu Academy of Agricultural Sciences,Key Laboratory of Animal Diseases Diagnostic and Immunology,Ministry of Agriculture,National Center for Engineering Research of Veterinary Bio-products,Nanjing 210014,China) |
| |
| Abstract: | The inosine 5-monophosphate dehydrogenase gene(impdh)was amplified by PCR from genome of Streptococcus suis 05ZYH33 isolate.The length of PCR fragment was 1 064 bp.The PCR product was digested with EcoR Ⅰ and Xho Ⅰ,and then was inserted into pET28a vector.The recombinant plasmid pET28a-impdh Was transformed into Escherichia coli BL21(DE3).After induced by 1 mmol/L IPTG,a high expressed protein in the E coli BL21 exhibited a band of 35 kD on SDS-PAGE.The expressed IMPDH showed good specific antigenicity and best enzymatic activity(reduced form of nicotinamide-adenine dinucleotid(NAD),NADH A_(340)=0.662～0.816)in optimal pH(pH 7.0～9.0)and at 37℃. |
| |
| Keywords: | Streptococcus suis serotype 2 inosine 5-monophosphate dehydrogenase(IMPDH) cloning enzymatic activity |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
|
