[1]张娜,张辉,胡圣伟,等.山羊痘病毒双向启动子序列的克隆与鉴定[J].中国预防兽医学报,2011,(08):585-588.
 ZHANG Na,ZHANG Hui,HU Sheng-wei,et al.Identification of a bidirectional promoter from Goatpox virus[J].Chinese journal of preventive veterinary medicine,2011,(08):585-588.
点击复制

山羊痘病毒双向启动子序列的克隆与鉴定
分享到:

《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2011年08期
页码:
585-588
栏目:
病原生物学
出版日期:
2011-08-15

文章信息/Info

Title:
Identification of a bidirectional promoter from Goatpox virus
作者:
张娜;张辉;胡圣伟;王安涛;张沾;陈创夫;乔军;
石河子大学生命科学学院动物科技学院;
Author(s):
ZHANG NaZHANG HuiHU Sheng-weiWANG An-taoZHANG ZhanCHEN Chuang-fu*QIAO Jun*
College of Life Science,College of Animal Science and Technology,Shihezi University,Shihezi 832000,China
关键词:
山羊痘病毒(GPV)双向启动子报告基因P56bPb56转录活性
Keywords:
Goatpox virus bidirectional promoter reporter gene promoter activity
摘要:
为鉴定山羊痘病毒(GPV)基因组中的启动子序列,本研究预测GPV基因组中早期转录因子VETF-l和中期转录因子VITF-3基因序列之间56 bp的一段序列为双向启动子,并将其命名为Pb56。通过PCR技术将该启动子序列的两个方向(Pb56+,Pb56-)分别与绿色荧光蛋白(EGFP)报告基因融合,构建重组转移重组质粒,分别命名为pUC-TK12-Pb56(+)-EGFP和pUC-TK12-Pb56(-)-EGFP。用脂质体转染法将2个转移重组质粒以及阴性对照pUC-TK12-EGFP及阳性对照pUC-TK12-P7.5-EGFP分别转染GPV预感染的BHK细胞;采用EGFP报告基因的表达水平评估双向启动子的活性。结果表明该基因序列的两个方向均能启动EGFP的表达,初步证实了该基因序列为GPV的双向启动子;Pb56+和Pb56-的转录活性均高于痘苗病毒的P7.5启动子。
Abstract:
In this study,a 56 bp sequence between ETF-l and VITF-3 genes in Goatpox virus(GPV) genome was predicted to be a bidirectional promoter,and the sequence was synthesized and verified for bidirection of promoting activites by insert the GFP gene under control on either side of the 56 bp sequence,respectively.The recombinant transfer plasmids of pUC-TK12-Pb56(+)-EGFP and pUC-TK12-Pb56(-)-EGFP were constructed based on GPV TK gene as homologous arms and transfected into BHK cells preinfected with GPV,respectively,with the positive control of pUC-TK12-P7.5-EGFP and negative control of pUC-TK12-EGFP.The results demonstrated that the promoting activites of the 56 bp sequence on both directions were stronger than the P7.5 of Vaccinia virus evaluated by the assay of EGFP reporter gene expression.The findings of the present studies showed that the 56 bp bidirection promotor had a potential use in construction of the recombinant virus vaccines based on GPV

参考文献/References:

[1] Matthews REF. Classification and nomenclature of viruses. Fourth Report of the International Committee on Taxonomy of Viruses. Intervirology. 1982
[2] Mackett M,Smith GL,Moss B. Vaccinia virus: a selectable eukaryotic cloning and expression vector. . 1982
[3] Zantinge J L,Krell P J,Derbyshire J B,et al. Partial transcriptional mapping of the fowlpox virus genome and analysis of the EcoRI L fragment. Journal of General Virology. 1996
[4] Romero C.H. Barrett t.Evans S.A.et.al. Single capripoxvirus recombinant vaccine for the protection of cattle against rinderpest and lumpy skin disease. Vaccine. 1993
[5] C.H. Romero,T. Barrett,R.W. Chamberlain,R.P. Kitching,M. Fleming,D.N. Black. Recombinant capripoxvirus expressing the haemagglutinin protein gene of rinderpest virus: protection of cattle against rinderpest and lumpy skin disease viruses. Journal of Virology. 1994
[6] Kumar S,Boyle D B. A poxvirus bidirectional promoter element with early/late and late functions. Journal of Virology. 1990
[7] Baldick C J,Jr,Keck J G,Moss B. Mutational analysis of thecore,spacer,and initiation regions of vaccinia virus intermedi-ate-class promoters[J]. Journal of Virology. 1992
[8] Wade-Evans A M,,Romero C H,Mellor P,et al.. Expression ofthe major core structural protein(VP7)of bluetongue virus by arecombinant capripox virus,provides partial protection of sheepagainst a virulent heterotypic bluetongue virus challenge[J].. Journal of Virology. 1996
[9] Cochran,MA,Puckett,C,Moss,B. In vitro mutagenesis of the promoter region for a vaccinia virus gene: evidence for tandem early and late regulatory signals. Journal of Virology. 1985
[10] Davian A J,Moss B. Structure of vaccinia virus early promoters. Journal of Molecular Biology. 1989
[11] 南文金,王清华,陆则基,陈飞,赵文姬,王志亮. 山羊痘病毒载体研究进展[J]. 动物医学进展. 2008年10期
[12] 邓伟,聂建超,唐祥,廖洪斌. 一起羊场暴发山羊痘的诊治[J]. 四川畜牧兽医. 2002年08期

备注/Memo

备注/Memo:
基金:国家973计划(2010CB30203);;国家自然科学基金(30800813)
更新日期/Last Update: 2011-09-29