ST细胞酵母双杂交cDNA文库的构建及鉴定  

Construction and Characterization of the Yeast Two-hybrid cDNA Library of ST Cells

作　　者：王鑫[1,2] 石达[2] 董慧[2] 曹丽艳[2] 陈建飞[2] 时洪艳[2] 张鑫[2] 冯力[1,2] WANG Xin SHI Da DONG Hui CAO Li-yan CHEN Jian-fei SHI Hong-yan ZHANG Xin FENG Li(College of Life Scicnce , Northcast Agricultural University, Harbin 150030, China Division of Swine Infectious Discases , State Key Laboratory of Veterinary Biotechnology , Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China)

机构地区：[1]东北农业大学生命科学学院,哈尔滨150030 [2]中国农业科学院哈尔滨兽医研究所,兽医生物技术国家重点实验室猪传染病研究室,哈尔滨150001

出　　处：《中国畜牧兽医》2017年第3期667-672,共6页China Animal Husbandry & Veterinary Medicine

基　　金：黑龙江省科研机构创新能力提升专项(YC2015D011);“十二五”农村领域国家科技计划课题(2015BAD12B02)

摘　　要：为了筛选与猪细小病毒(porcine parvovirus,PPV)结构蛋白VP2相互作用的宿主蛋白,深入研究病毒入侵及致病的分子机制,本试验建立了ST细胞酵母双杂交cDNA文库。采用RNeasy Min Kit提取ST细胞的总RNA,反转录合成cDNA第一链之后,通过SMART技术合成了双链cDNA,并利用同源重组的方法,构建了ST细胞的酵母cDNA文库。结果显示,文库滴度为8.1×108 CFU/mL,插入的双链cDNA片段大小为250~1 000bp,平均大小约为500bp,文库的重组率为96%。此文库可用于筛选与病毒相互作用的ST细胞宿主蛋白,为进一步阐明病毒的致病机制奠定了基础。To seek out proteins interact with VP2 structural protein of porcine parvovirus（PPV）in ST cell and study the molecular mechanisms of viral infection and pathogenicity,a ST cDNA yeast hybrid library was created.Firtly,the total RNA of ST was extracted using RNeasy Min Kit,and double strands cDNA was synthesized by SMART technique.Then,the ST cDNA library was successfully created by homologous recombination in yeast,the titer was 8.1×10^8 CFU/mL,the inserts varied from 250 to 1 000 bp,the average of inserted fragments was about 500 bp,and the recombination rate was 96%.These datas indicated that the yeast twohybrid cDNA library was successfully constructed and might be useful for screening the host proteins interacting with structure protein of PPV.

关 键 词：猪细小病毒 酵母双杂交 ST细胞 CDNA文库 

分 类 号：Q782[生物学—分子生物学]