中国兽医科学2012，Vol.42Issue(5)：454-460,7.
新型鸡痘病毒穿梭载体的构建及其体外表达
Construction and expression in vitrostudies on shuttle vector of fowlpox virus
摘要
Abstract
To construct a new vector for screening recombinant fowlpox virus,thymidine kinase（TK） gene of fowlpox virus（FPV） 282E4 strain was partial sequenced using chromosome walking techniques.Expression cassettes containing promoters,multiple cloning site（MCS） and termination signals were designed and synthesized so as to express three exogenous genes.The expression cassettes were inserted between the two arms of the TKL（TK left） and TKR（TK right） gene,and then the new fowlpox virus shuttle vector pTKE3 was constructed.To verify the features of expression cassette,the enhanced green fluorescent protein gene was cloned into downstream of three promoter to construct three plasmids.The constructed plasmids pTKE3-A,pTKE3-B and pTKE3-C which were transfected by lipofectamine on chicken embryo fibroblasts cell pre-infected with FPV 282E4 respectively,and the expression efficiency was identified by observing fluorescence after transfection at 12 h,which show that the shuttle vector containing three cassettes can express foreign proteins successfully.The present study lays a foundation for further construction of a polyvalent live vector vaccine of recombinant fowlpox virus.关键词
鸡痘病毒/穿梭载体/表达Key words
fowlpox virus/shuttle vector/expression分类
农业科技引用本文复制引用
刘存霞,李昌,王茂鹏,胡乐鹏,靖杰,杜寿文,尹荣兰,刘燕瑜,任静强,任大勇,孙丹丹,金宁一..新型鸡痘病毒穿梭载体的构建及其体外表达[J].中国兽医科学,2012,42(5):454-460,7.基金项目
国家自然科学基金项目 （）
国家重点基础研究发展计划（973）项目前期研究专项 （）
吉林省中青年科技领军人才及优秀创新团队项目 （）
吉林省高新技术产业发展项目 （）
