Suicide Vector Construction of Haemophilus parasuis hhd B Gene Marker-free Deleted  

作　　者：Song Shuai Li Miao Li Yan Jiang Zhiyong Cai Rujian Yang Dongxia Li Chunling 

机构地区：[1]Institute of Animal Health,Guangdong Academy of Agricultural Sciences , Guangdong Key Labratory of Animal Disease Prevention

出　　处：《Animal Husbandry and Feed Science》2015年第6期354-357,共4页动物与饲料科学（英文版）

基　　金：Supported by Science and Technology Project of Guangdong Province(2013B020307002)

摘　　要：To construct the suicide vector of hhd B gene marker-free mutant in Haemophilus parasuis( HPS),two pairs of specific primers were designed and synthesized according to the hhd B gene upstream and downstream sequences of HPS published in Gen Bank. The hhd B gene upstream and downstream sequences were amplified by PCR,which were further ligated( hhd B-up + down) through overlapping PCR method. NotⅠand SalⅠrestriction enzyme sites were introduced on both ends of the ligated sequence. After the corresponding digestion,the hhd B-up + down sequence was directionally cloned to the suicide plasmid vector p EMOC2. Results showed that the suicide vector of hhd B gene marker-free deleted( p EMOC2Δhhd B) with stable inheritance in E. coli β2155 strain was successfully obtained,thereby laying the foundation for construction of HPS-hhd B gene marker-free mutant strain.To construct the suicide vector of hhd B gene marker-free mutant in Haemophilus parasuis( HPS),two pairs of specific primers were designed and synthesized according to the hhd B gene upstream and downstream sequences of HPS published in Gen Bank. The hhd B gene upstream and downstream sequences were amplified by PCR,which were further ligated( hhd B-up + down) through overlapping PCR method. NotⅠand SalⅠrestriction enzyme sites were introduced on both ends of the ligated sequence. After the corresponding digestion,the hhd B-up + down sequence was directionally cloned to the suicide plasmid vector p EMOC2. Results showed that the suicide vector of hhd B gene marker-free deleted( p EMOC2Δhhd B) with stable inheritance in E. coli β2155 strain was successfully obtained,thereby laying the foundation for construction of HPS-hhd B gene marker-free mutant strain.

关 键 词：Haemophilus parasuis(HPS) hhd B gene Suicide vector 

分 类 号：S852.61[农业科学—基础兽医学]