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| 表达绵羊肺腺瘤病毒跨膜蛋白HepG2细胞系的建立 | |
| 引用本文： | 刘霄卉,罗军荣,斯日古楞,周建华,马学恩. 表达绵羊肺腺瘤病毒跨膜蛋白HepG2细胞系的建立[J]. 中国预防兽医学报, 2011, 33(4). DOI: 10.3969/j.issn.1008-0589.2011.04.18 |
| 作者姓名： | 刘霄卉 罗军荣 斯日古楞 周建华 马学恩 |
| 作者单位： | 1. 内蒙古农业大学兽医学院,内蒙古呼和浩特,010018 2. 内蒙古农业大学兽医学院,内蒙古呼和浩特010018;江西农业大学动物科学技术学院,江西南昌330045 3. 中国农业科学院哈尔滨兽医研究所,黑龙江哈尔滨,150001 |
| 基金项目： | 国家自然科学基金，兽医生物技术国家重点实验室开放课题基金 |
| 摘 要： | 为深入研究绵羊肺腺瘤病毒(JSRV)与绵羊肺腺瘤病(OPA)发病关系,本研究采用PCR方法从pGEX-4T-1-TM重组质粒中扩增编码JSRV跨膜蛋白(TM)的基因序列,并引入标签多肽HA序列和限制性内切酶位点,将其重组至真核表达载体pcDNA3.1(+)中,构建了重组质粒pcDNA-TM-HA.通过转染HepG2细胞并用G418筛选,对稳定表达TM的阳性细胞进行纯化,获得了稳定表达JSRV tm基因的HepG2细胞系.间接免疫荧光及western blot检测结果表明,重组蛋白TM-HA在HepG2细胞中得到正确表达.JSRV TM蛋白稳定表达细胞系的建立为进一步研究该蛋白与JSRV诱导OPA发病关系提供了重要的实验平台. |
| 关 键 词： | 绵羊肺腺瘤反转录病毒 跨膜蛋白 真核表达 HepG2细胞系 |
Development of a HepG2 cell line expressing the transmembrane protein of Jaagsiekte retrovirus |
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| LIU Xiao-hui,LUO Jun-rong,Siriguleng,ZHOU Jian-hua,MA Xue-en. Development of a HepG2 cell line expressing the transmembrane protein of Jaagsiekte retrovirus[J]. Chinese Journal of Preventive Veterinary Medicine, 2011, 33(4). DOI: 10.3969/j.issn.1008-0589.2011.04.18 | |
| Authors: | LIU Xiao-hui LUO Jun-rong Siriguleng ZHOU Jian-hua MA Xue-en |
| Affiliation: | LIU Xiao-hui1,LUO Jun-rong1,2,Siriguleng1,ZHOU Jian-hua3*,MA Xue-en1* (1. College of Veterinary Science,Inner Mongolia Agricultural University,Huhhot 010018,China,2. College of Animal Science and Technology,Jiangxi Agricultural University,Nanchang 330045,3. Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150001,China) |
| Abstract: | To study the relationship between jaagsiekte retrovirus (JSRV) transmembrane protein (TM) and the pathogenesis of ovine pulmonary adenomatosis (OPA), a HepG2 cell line expressing the TM of JSRV was developed. The coding region of tm gene was amplified with an HA tag by PCR, and was cloned into the eukaryotic expression vector pcDNA3.1(+). The resulting recombinant plasmid was transfected into HepG2 cells and selected under the G418. A positive cells which stable expression of JSRV TM was selected and purifi... |
| Keywords: | jaagsiekte sheep retrovirus transmambrane protein eukaryotic expression HepG2 cell line |
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