[1]高兴红,贾仁勇*.鸭肠炎病毒 UL24/C基因酵母双杂交诱饵载体构建及鉴定[J].中国预防兽医学报,2018,(06):481-485.[doi:0.3969/j.issn.1008-0589.201709036]
 GAO Xing-hong,JIA Ren-yong*.Construction and identification bait vector of duck enteritis virus UL24/C gene in yeast two-hybrid system[J].Chinese journal of preventive veterinary medicine,2018,(06):481-485.[doi:0.3969/j.issn.1008-0589.201709036]
点击复制

鸭肠炎病毒 UL24/C基因酵母双杂交诱饵载体构建及鉴定()
分享到:

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

卷:
期数:
2018年06
页码:
481-485
栏目:
病原生物学
出版日期:
2018-06-20

文章信息/Info

Title:
Construction and identification bait vector of duck enteritis virus UL24/C gene in yeast two-hybrid system
文章编号:
1008-0589(2018)6-0481-05
作者:
 

高兴红123贾仁勇234*

 (1. 遵义医学院 贵州省普通高等学校传染病与生物安全特色重点实验室,贵州 遵义 563000;
2. 四川农业大学 禽病防治研究中心,四川 成都 611130;3. 四川农业大学 预防兽医研究所,四川 成都 611130;
4. 动物疫病与人类健康四川省重点实验室,四川 成都 611130)
Author(s):
 

GAO Xing-hong123 JIA Ren-yong234*

 (1. Key Laboratory of Infectious Diseases and Biosecurity, Provincial Department of Education, Zunyi Medical College, Zunyi 563000, China; 2. Avian Disease Research Center, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China;
3. Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China;
4. Key Laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu 611130, China)
关键词:
鸭肠炎病毒UL24基因酵母双杂交诱饵载体
Keywords:
duck enteritis virus UL24 gene yeast teo-hybrid bait vector
分类号:
S852.65
DOI:
0.3969/j.issn.1008-0589.201709036
文献标志码:
A
摘要:
为构建DEV UL24/C蛋白的酵母双杂交的诱饵载体,本研究以DEV CHv株基因组DNA为模板,经一步法克隆构建pGBKT7-UL24/C诱饵质粒,质粒转化酵母菌株后进行自激活活性检测、免疫印迹检测和毒性检测。结果显示,诱饵质粒pGBKT7-UL24/C插入片段大小为510 bp,且未发生基因突变;pGBKT7-UL24/C质粒转化Y2HGold菌株后的自激活检测显示诱饵菌株无自激活活性,免疫印迹检测表明该诱饵菌株成功表达DEV UL24/C蛋白,毒性检测表明该诱饵蛋白对宿主菌Y2HGold无毒性作用。综上表明,未研究正确构建了UL24/C基因酵母双杂交诱饵载体Y2HGold (pGBKT7-UL24/C),为筛选与DEV UL24蛋白相互作用蛋白、探究DEV UL24蛋白功能及分子机制奠定了基础。
  
Abstract:
To generate a bait strain Y2HGold (pGBKT7-UL24/C) using yeast two-hybrid, the bait plasmid of pGBKT7- UL24/C was constructed by one-step cloning with UL24/C gene fragment amplified from genomic DNA of duck enteritis virus (DEV) CHv strain as a template. And after the plasmid being transformed into yeast strain, the bait protein was detected by self- activating activity, western blot and toxicity assays. The results indicated that the insert fragment in bait plasmid of pGBKT7- UL24/C was 510bp. Self-activation assay revealed that bait protein had no self-activation activity when the pGBKT7-UL24/C plasmid was transformed to Y2HGold strain. Western blot analysis further showed DEV UL24/C protein was expressed successfully in the bait strain. And the toxicity test showed that bait protein had no toxic to the host strain of Y2HGold. In summary, the yeast two-hybrid bait vector of Y2HGold (pGBKT7-UL24/C) was successfully constructed. And this study also layed the foundation for screening proteins interaction with DEV UL24 and for exploring the function and for exploring the function and molecular mechanism of DEV UL24 protein.

参考文献/References:

 
[1]King A M Q, Lefkowitz E, Adams M J, et al. Virus Taxonomy: tenth Report of the International Committee on Taxonomy of Viruses [M]. Elsevier: Academic Press, 2016.
[2]Li Yu-feng, Huang Bing, Ma Xiu-li, et al. Molecular characterization of the genome of duck enteritis virus [J]. Virology, 2009, 391(2): 151-161.
[3]Wang Ji-chun, Dirk H, Martin B, et al. Complete genome sequence of virulent duck enteritis virus (DEV) strain 2085 and comparison with genome sequences of virulent and attenuated DEV strains [J]. Virus Res, 2011, 160(1): 316-325.
[4]Wu Ying, Cheng An-chun, Wang Ming-shu, et al. Complete genomic sequence of chinese virulent duck enteritis virus [J]. J Virol, 2012, 86(10): 5965-5965.
[5]Wu Ying, Cheng An-chun, Wang Ming-shu, et al. Comparative genomic analysis of duck enteritis virus strains [J]. J Virol, 2012, 86(24): 13841-13842.
[6]Susan B, Jacek K, Donna T, et al. Herpes simplex virus type 2 UL24 gene is a virulence determinant in murine and guinea pig disease models [J]. J Virol, 2005, 79(16): 10498-10506.
[7]Luc B, Gabriel Andre L T, Huda H, et al. Conserved residues in the UL24 protein of herpes simplex virus 1 are important for dispersal of the nucleolar protein nucleolin [J]. J Virol, 2010, 84(1): 109-118.
[8]Xu Hai-yan, Su Chen-he, Angela. P, et al. Herpes simplex virus 1 Ul24 abrogates the DNA sensing signal pathway by inhibiting Nf-Κb activation [J]. J Virol, 2017, 91(7): e00025-00017.
[9]Lin S R, Jiang M J, Wang H H, et al. Human cytomegalovirus UL76 elicits novel aggresome formation via interaction with S5a of ubiquitin proteasome system [J]. J  Virol, 2013, 87(21): 11562-11578.
[10]Helena C, Rute N, John S, et al. Human cytomegalovirus gene UL76 induces Il-8 expression through activation of the DNA damage response [J]. PLoS pathog, 2013, 9(9): e1003609.
[11]Weller S K, Schumacher A J, Mohni K N. Herpes simplex virus: Manipulating DNA damage response pathways [J]. FASEB J, 2012, 26: 932.
[12]Nascimento R, Dias J D, Parkhouse R M. The conserved UL24 family of human alpha, beta and gamma herpesviruses induces cell cycle arrest and inactivation of the cyclinb/Cdc2 complex [J]. Arch Virol, 2009, 154(7): 1143-1149.
[13]Yu Xia, Jia Ren-yong, Huang Juan, et al. Attenuated Salmonella typhimurium delivering DNA vaccine encoding duck enteritis virus UL24 induced systemic and mucosal immune responses and conferred good protection against challenge [J]. Vet Res, 2012, 43(1): 1-10.
[14]Jia Ren-yong, Cheng An-chun, Wang Ming-shu, et al. Cloning, expression, purification and characterization of UL24 partial protein of duck enteritis virus [J]. Intervirology, 2009, 52(6): 326-334.
[15]Jia Ren-yong, Cheng An-chun, Wang Ming-shu, et al. Development and evaluation of an antigen-capture elisa for detection of the UL24 antigen of the duck enteritis virus, based on a polyclonal antibody against the UL24 expression protein [J]. J Virol Methods, 2009, 161(1): 38-43.
[16]Liu Xue-yan, Liu Qing, Xiao Kang-peng, et al. Attenuated Salmonella typhimurium delivery of a novel DNA vaccine induces immune responses and provides protection against duck enteritis virus [J]. Vet Microbiol, 2016, 186: 189-198.
[17]Tu Jian-feng, Xu Jia-ping, Liu Hui-tao, et al. Determination and analysis of complete mitochondrial genome sequence of peking duck (Anas platyrhychos) [J]. Mitochondrial DNA Part A, 2016, 27(1): 682-683.
[18]Zhang Guo-jie, Li Cai, Li Qi-ye, et al. Comparative genomics reveals insights into avian genome evolution and adaptation [J]. Science, 2014, 346(6215): 1311-1320.
[19]Gao Xing-hong, Jia Ren-yong, Wang Ming-shu, et al. Duck enteritis virus (DEV) UL54 protein, a novel partner, interacts with DEV UL24 protein [J]. Virol J, 2017, 14(1): 166.
[20]Li Qiao, Chen Pan, Zeng Zhao-yang, et al. Yeast two-hybrid screening identified WDR77 as a novel interacting partner of TSC22D2 [J]. Tumor Biology, 2016, 37(9): 12503-12512.
                                        (本文编辑:彭永刚)

相似文献/References:

[1]王克雄,李慧昕,李 阳,等.鸭肠炎病毒VP22蛋白单克隆抗体的制备及鉴定 [J].中国预防兽医学报,2009,(09):721.
 WANG Ke-xiong,LI Hui-xin,et al.Preparation and identification of monoclonal antibodies against VP22 protein of duck enteritis virus [J].Chinese journal of preventive veterinary medicine,2009,(06):721.
[2]陈淑红,刘怀然,韩宗玺,等.应用PCR检测鸭肠炎病毒弱毒在鸡胚体内的分布[J].中国预防兽医学报,2007,(01):17.
 CHEN Shu-hong,LIU Huai-ran,HAN Zong-xi,et al.Detection of the distribution of duck enteritis virus attenuated strain in chicken embryos by using PCR[J].Chinese journal of preventive veterinary medicine,2007,(06):17.
[3]李 冰,李慧昕,王 钰,等.鸭肠炎病毒gL蛋白在鸡胚成纤维细胞中动态表达的检测[J].中国预防兽医学报,2012,(01):19.[doi:10.3969/j.issn.1008-0589.2012.01.05]
 LI Bing,LI Hui-xin,WANG Yu,et al.Detection of the duck enteritis virus gL protein expressed in chicken embryo fibroblast[J].Chinese journal of preventive veterinary medicine,2012,(06):19.[doi:10.3969/j.issn.1008-0589.2012.01.05]
[4]王玉龙,李慧昕,李 冰,等.TK基因缺失对鸭肠炎病毒在鸡胚成纤维细胞中复制影响的评价[J].中国预防兽医学报,2015,(05):339.[doi:10.3969/j.issn.1008-0589.2015.05.04]
 WANG Yu-long,LI Hui-xin,LI Bing,et al.Evaluation on the effect of duck enteritis virus replicationin CEF influenced by the deletion of TK gene[J].Chinese journal of preventive veterinary medicine,2015,(06):339.[doi:10.3969/j.issn.1008-0589.2015.05.04]
[5]张苗苗,李慧昕,韩宗玺,等.表达新城疫病毒F基因重组鸭肠炎病毒的构建[J].中国预防兽医学报,2016,(02):92.[doi:10.3969/j.issn.1008-0589.2016.02.02]
 ZHANG Miao-miao,LI Hui-xin,HAN Zong-xi,et al.Construction of recombinant duck enteritis virus expressing F gene of Newcastle disease virus[J].Chinese journal of preventive veterinary medicine,2016,(06):92.[doi:10.3969/j.issn.1008-0589.2016.02.02]

更新日期/Last Update: 2018-07-11