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| 细粒棘球蚴SmadC蛋白及其MH2结构域酵母双杂交载体的构建及自激活鉴定 | |
| 引用本文： | 李静,张传山,吕国栋,王俊华,魏绪法,李朝旺,范金亮,林仁勇,剡根强.细粒棘球蚴SmadC蛋白及其MH2结构域酵母双杂交载体的构建及自激活鉴定[J].中国兽医学报,2012,32(1):58-62. |
| 作者姓名： | 李静 张传山 吕国栋 王俊华 魏绪法 李朝旺 范金亮 林仁勇 剡根强 |
| 作者单位： | 1. 石河子大学动物科技学院,新疆石河子832003／新疆医科大学第一附属医院医学研究中心新疆包虫病基础医学重点实验室,新疆乌鲁木齐830054 2. 新疆医科大学第一附属医院医学研究中心新疆包虫病基础医学重点实验室,新疆乌鲁木齐,830054 3. 石河子大学动物科技学院,新疆石河子,832003 |
| 基金项目： | 国家自然科学基金资助项目(30860253);新疆维吾尔自治区包虫病基础医学重点实验室开放课题(XJDX0202-2010-06) |
| 摘 要： | 从重组克隆载体pMD18-T—egsmadC中扩增细粒棘球蚴egsmadC基因及其MH2编码序列，将其克隆入酵母双杂交载体pGADT7和pGBKT7中，经PCR、限制性酶切鉴定及序列测定正确后，PEG／LiAe法转化酵母菌株，测定融合蛋白对酵母菌生长的影响和自激活活性。结果表明，egsmadC基因及其MH2编码序列表达的蛋白对酵母茵Y2HGold无毒性和自激活作用，可为进一步运用酵母双杂交技术筛选与之相互作用的蛋白奠定基础。 |
| 关 键 词： | smadC基因 MH2结构域 酵母双杂交 自激活 |
Construction of DNA-AD and DNA-BD vectors with EgSmadC and MH2 domain for yeast two-hybrid system |
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| LI Jing,ZHANG Chuan-shan,LU Guo-dong,WANG Jun-hua,WEI Xu-fa,LI Chao-wang,FAN Jin-liang,LIN Ren-yong,YAN Gen-qiang.Construction of DNA-AD and DNA-BD vectors with EgSmadC and MH2 domain for yeast two-hybrid system[J].Chinese Journal of Veterinary Science,2012,32(1):58-62. | |
| Authors: | LI Jing ZHANG Chuan-shan LU Guo-dong WANG Jun-hua WEI Xu-fa LI Chao-wang FAN Jin-liang LIN Ren-yong YAN Gen-qiang |
| Institution: | 1*(1.College of Animal Science &Technology,Shihezi University,Shihezi,Xinjiang 832003,China;2.Medical Research Center and Xinjiang Key Lab or Rtory of Echinococcosis,The First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,China) |
| Abstract: | The complete encoding gene of egsmadC and egsmadC-MH2 from Echinococcos granulosus(Eg) was amplified in the recombinant plasmid pMD18-T-egsmadC by PCR and they were then subcloned into MCS of pGADT7 and pGBKT7 vectors,respectively.After being verified by PCR,restriction analysis and sequencing,the recombined plasmids were transformed into the yeast strain Y2HGold by PEG/LiAc method and their toxicity and self-activation were detected.This result means that the yeast two-hybrid expression vectors of egsmadC and egsmadC-MH2 were constructed successfully and would not toxicity and self-activation,which lays the foundations for screening the egsmadC-interacting proteins using the yeast two-hybrid technique. |
| Keywords: | smadC gene MH2 domain yeast two-hybrid system self-activation |
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