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| 弓形虫ROP2基因的克隆及原核表达 | |
| 引用本文： | 江涛,周艳琴,刘琴,聂浩,姚宝安,赵俊龙.弓形虫ROP2基因的克隆及原核表达[J].中国兽医学报,2007,27(4):527-531. |
| 作者姓名： | 江涛 周艳琴 刘琴 聂浩 姚宝安 赵俊龙 |
| 作者单位： | 1. 华中农业大学农业微生物学国家重点实验室,湖北,武汉,430070;华中农业大学动物医学院,湖北,武汉,430070;长江大学动物科学学院,湖北,荆州,434100 2. 华中农业大学动物医学院,湖北,武汉,430070 3. 华中农业大学农业微生物学国家重点实验室,湖北,武汉,430070;华中农业大学动物医学院,湖北,武汉,430070 |
| 基金项目： | 高等学校博士学科点专项科研项目;教育部科学技术研究项目 |
| 摘 要： | 应用PCR技术从刚地弓形虫（Toxoplasma gondii）RH株的基因组DNA中扩增编码棒状体蛋白ROP2（rh—potryprotein2）的部分基因，构建pGEX—KG—ROP2重组表达质粒，经酶切、PCR及DNA测序鉴定后，将阳性质粒转入E．coli BL21CodonPlus中，在IPTG诱导下表达，表达产物用SDS—PAGE和Western—blot分析鉴定。结果表明，扩增的ROP2基因与GenBank上发表的相应基因序列的同源性达99.9％，该基因可以在大肠杆菌中高效表达，表达的ROP2融合蛋白表观相对分子质量约为64000，可被兔抗弓形虫免疫血清识别。 |
| 关 键 词： | 刚地弓形虫 棒状体蛋白ROP2 基因克隆 原核表达 |
| 文章编号： | 1005-4545（2007）04-0527-05 |
| 修稿时间： | 2005-09-23 |
Cloning and protokaryotic expression of the Toxoplasma gondii ROP2 gene |
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| JIANG Tao,ZHOU Yan-qin,LIU Qin,NIE Hao,YAO Bao-an,ZHAO Jun-long.Cloning and protokaryotic expression of the Toxoplasma gondii ROP2 gene[J].Chinese Journal of Veterinary Science,2007,27(4):527-531. | |
| Authors: | JIANG Tao ZHOU Yan-qin LIU Qin NIE Hao YAO Bao-an ZHAO Jun-long |
| Institution: | 1. National Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University , Wuhan 430070, China;2. College of Animal Veterinary Medicine, Huazhong Agricultural University , Wuhan 430070, China ; 3. College of Animal Science,Yangtze University,Jingzhou,Hubei 434100, China |
| Abstract: | The truncated ROP2 gene was obtained by amplification from genomic DNA of Toxoplasma gondii RH strain and cloned into the plasmid pGEX-KG, The constructed recombinant plasmid was identified by enzyme digesting, PCR and sequencing , Then the target recombinant plasmid was transferred into E, coli BL21Codonplus susceptible cell, the expression of the recombinant pGEX-KG-ROP2 was induced by IPTG, The expression protein were analysed by SDS-PAGE and by Western-blotting, The results showed that the sequence identities are 99, 9 % between amplified ROP2 gene and that from GenBank, The molecular weight of fused ROP2 was about 64 000 by SDS-PAGE. It could be specifically recognized by immune serum against Toxoplasma gondii from rabbit. |
| Keywords: | Toxoplasma gondii rhoptry protein ROP2 gene clone protokaryotic expression |
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