[1]刘霄卉,罗军荣,斯日古楞,等.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,(04):323-326.
 LIU Xiao-hui,LUO Jun-rong,Siriguleng,et al.Development of a HepG2 cell line expressing the transmembrane protein of Jaagsiekte retrovirus[J].Chinese journal of preventive veterinary medicine,2011,(04):323-326.
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表达绵羊肺腺瘤病毒跨膜蛋白HepG2细胞系的建立
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2011年04期
页码:
323-326
栏目:
研究简报
出版日期:
2011-04-15

文章信息/Info

Title:
Development of a HepG2 cell line expressing the transmembrane protein of Jaagsiekte retrovirus
作者:
刘霄卉;罗军荣;斯日古楞;周建华;马学恩;
内蒙古农业大学兽医学院;江西农业大学动物科学技术学院;中国农业科学院哈尔滨兽医研究所;
Author(s):
LIU Xiao-hui1 LUO Jun-rong12 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, China; 3. Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China
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
Keywords:
jaagsiekte sheep retrovirus transmambrane protein eukaryotic expression HepG2 cell line
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。
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 purified. The cells expression of JSRV TM was confirmed by indirect immunofluorescence assay and western blot. This cell line provides a useful platform for studies on JSRV in vitro

参考文献/References:

[1] Bai J,Bishop JV,Carlson JO,DeMartini JC. Sequence comparison of JSRV with endogenous proviruses:envelope genotypes ang a novel ORF with similarity to a G-protein-coupled receptor. Journal of Virology. 1999
[2] Caporale M,Arnaud F,Mura M,Golder M,Murgia C,Palmarini M. The Signal Peptide of a Simple Retrovirus Envelope Functions as a Posttranscriptional Regulator of Viral Gene Expression. Journal of Virology. 2009
[3] Alberti A,Murgia C,et al. Envelope—induced transformation by ovine betaretroviruses. Journal of Virology. 2002
[4] Rous P. A sarcoma of the fowl transmissible by an agent separable from the tumor cells. The Journal of Experimental Medicine. 1911
[5] Liu Shan-lu,Miller A D. Oncogenic transformation by the jaag-siekte sheep retrovirus envelope protein[J]. Oncegene. 2007
[6] Griffiths D J,Martineau H M,Cousens C. Pathology and patho-genesis of ovine pulmonary adenocarcinoma[J]. Journal of Comparative Pathology. 2010
[7] Cousens C,Maeda N,Murgia C,et al. In vivo tumorigenesis by Jaagsiekte sheep retrovirus (JSRV)requires Y590in Env TM,but not full-length orfX open reading frame[J]. Journal of Virology. 2007
[8] 斯日古楞,么宏强,马学恩,王升启. 绵羊肺腺瘤病毒内蒙毒株囊膜蛋白基因在大肠杆菌中的分段表达[J]. 黑龙江畜牧兽医. 2007年10期

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备注/Memo

备注/Memo:
基金:国家自然科学基金(31060332);;兽医生物技术国家重点实验室开放课题基金(SKLVBF2008)
更新日期/Last Update: 2011-09-29