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| 本地毛形线虫49 Ku ES蛋白结构基因的分子克隆及原核表达 | |
| 引用本文： | 郑宝亮,王秀荣,路义鑫,李冬梅,闫清波,宋铭忻.本地毛形线虫49 Ku ES蛋白结构基因的分子克隆及原核表达[J].中国预防兽医学报,2007,29(7):510-514. |
| 作者姓名： | 郑宝亮 王秀荣 路义鑫 李冬梅 闫清波 宋铭忻 |
| 作者单位： | 1. 郑州牧业工程高等专科学校,生物工程系,河南,郑州,450011 2. 中国农业科学院哈尔滨兽医研究所,农业部动物流感重点开放实验室/兽医生物技术国家重点实验室,黑龙江,哈尔滨,150001 3. 东北农业大学动物医学院预防医学系,寄生虫学教研室,黑龙江,哈尔滨,150030 |
| 基金项目： | 致谢：本研究大部分是在中国农业科学院哈尔滨兽医研究所农业部动物流感重点开放实验室以及兽医生物技术国家重点实验室进行和完成的,对实验室在技术上的指导与实验用品及使用仪器方面的支持表示感谢. |
| 摘 要： | 提取Trichinella nativa(T.nativa)肌幼虫的总RNA,用RT-PCR方法扩增出了编码T.nativa 49 Ku ES蛋白的结构基因。基因克隆后测序,序列测定结果表明:目的基因TNPG长度为951 bp,核苷酸序列同已发表的Trichinella spiralis(T.spiralis)相应的序列P49同源性为97.68%,所推导的氨基酸序列同源性为95.24%。将目的基因TNPG插入到原核表达载体pET-30a的BamHⅠ酶切位点处,并转化到感受态表达菌中进行诱导表达。结果显示TNPG在原核表达菌BL-21中获得了高效表达,表达产物为40.8 Ku的融合蛋白,表达量达到菌体总蛋白的22.8%。通过Western blot分析,表达产物可以被小鼠T.nativa和T.spiralis阳性血清以及它们的中国地理株的小鼠血清特异性识别。 |
| 关 键 词： | 旋毛虫nativa种 旋毛虫spiralis种 49 Ku Es蛋白 基因序列 Western blot分析 |
| 文章编号： | 1008-0589（2007）07-0510-05 |
| 修稿时间： | 2006-11-28 |
Cloning and prokaryotic expression of 49 Ku ES protein structural gene for Trichinella nativa |
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| ZHENG Bao-liang,WANG Xiu-rong,LU Yi-xin,LI Dong-mei,YAN Qing-bo,SONG Ming-xin.Cloning and prokaryotic expression of 49 Ku ES protein structural gene for Trichinella nativa[J].Chinese Journal of Preventive Veterinary Medicine,2007,29(7):510-514. | |
| Authors: | ZHENG Bao-liang WANG Xiu-rong LU Yi-xin LI Dong-mei YAN Qing-bo SONG Ming-xin |
| Institution: | 1. Bioengineering Department, Zhengzhou College of Animal Husbandry and Engineering, Zhengzhou 450011, China; 2. National Key Laboratory of Veterinary Biotechnology, Avian Influenza Lab of Ministry of Agriculture, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 150001, China; 3. Section of Parasitology, Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China |
| Abstract: | The gene TNPG encoding the 49 Ku ES protein of Trichinella nativa(T.nativa)was amplified by RT-PCR and sequenced.The gene consisted of 951 bp and shared 97.68 % nucleotide homology with the sequence of P49 from Trichinella spiralis (T.spiralis),and 95.24 % homology of deduced amino acid between TNPG and P49.TNPG was cloned into the prokaryotic expression vector pET-30a and expressed in E.coli cells.The expressed product was 40.8 Ku in size and accounted for 22.8 % of the total protein.Western blot analysis showed that the expressed product reacted with sera of mice infected with T.nativa,T.spiralis and other geographical isolates in China. |
| Keywords: | Trichinella nativa Trichinella spiralis 49 Ku ES protein gene sequence Western blot analysis |
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