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扩展莫尼茨绦虫烯醇化酶基因的原核表达及抗原性鉴定
引用本文:夏党荣,陈南颖,马勋,薄新文,何延华,康立超.扩展莫尼茨绦虫烯醇化酶基因的原核表达及抗原性鉴定[J].中国预防兽医学报,2012,34(3):241-243,250.
作者姓名:夏党荣  陈南颖  马勋  薄新文  何延华  康立超
作者单位:1. 石河子大学动物科技学院,新疆石河子832000;新疆农垦科学院新疆兵团绵羊繁育生物技术重点实验室,新疆石河子832000
2. 石河子大学动物科技学院,新疆石河子,832000
3. 新疆农垦科学院新疆兵团绵羊繁育生物技术重点实验室,新疆石河子,832000
基金项目:国家重点基础研究发展计划(973)前期研究专项(2006CB708512);国家绒毛用羊产业技术体系建设专项(CARS-40-11);兰兽研国家重点实验室开放课题(SKLVEB2011KFKT008);兵团杰出青年科学基金(2011CD005)
摘    要:为表达扩展莫尼茨绦虫(Moniezia expansa)的烯醇化酶(Enolase),本研究采用RT-PCR方法以M.expansa 组织总RNA为模板扩增enolase基因,将其克隆到原核表达栽体pET-28a中,并转化BL21 (DE3)感受态细胞,以IPTG进行诱导表达.SDS-PAGE结果显示表达的重组Enolase约为47 ku.Western blot分析表明,天然蛋白能够被重组表达产物免疫小鼠的血清识别,出现两条免疫印记条带,即天然蛋白以两种形式(约47 ku和40 ku)存在,并具有抗原性,为该蛋白功能研究奠定了基础.

关 键 词:扩展莫尼茨绦虫  烯醇化酶基因  原核表达  抗原性

Cloning and prokaryotic expression and antigenicity identification of enolase from Moniezia expansa
XIA Dang-rong , CHEN Nan-ying , MA Xun , BO Xin-wen , HE Yan-hua , KANG Li-chao.Cloning and prokaryotic expression and antigenicity identification of enolase from Moniezia expansa[J].Chinese Journal of Preventive Veterinary Medicine,2012,34(3):241-243,250.
Authors:XIA Dang-rong  CHEN Nan-ying  MA Xun  BO Xin-wen  HE Yan-hua  KANG Li-chao
Institution:1.College of Animal Science and Technology,Shihezi University,Shihezi 832000,China;2.Xinjiang Production and Construction Corps,Key Laboratory of Ovine Breed and Biotechnology,Xinjiang Academy of Agricultural and Reclamation,Shihezi 832000,China)
Abstract:To clone and express the coding enolase gene from Moniezia expansa.One pair of specific primers was designed according to M.expansa enolase cDNA in Genbank(JF803807).Total RNA was extracted from M.expansa and the enolase gene was amplified by RT-PCR and cloned into pET28a vector.The recombinant plasmid pET28a-Enolase was transformed into E.coli BL21(DE3) and induced with IPTG for expression.The results showed that the recombinant enolase was about 47 ku and two protein bands(about 47 ku and 40 ku) were detected in the natural enolase with positive serum prepured from mice.Those results would be useful to further study of the enolase function immuned with the recombinant protein.This is the base of researching into enolase gene function and genetic engineering application.
Keywords:Moniezia expansa  enolase gene  prokaryotic expression  antigenicity
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