布氏杆菌抗原蛋白pbp39基因重组穿梭质粒的构建与鉴定  

Construction and Identification of Recombinant Shuttle-plasmid with pbp39 from Brucella

作　　者：钟蓓[1] 杨玉莹[1] 张西臣[2] 顾玉芳[1] 陈振威[1] 多婷[1] 

机构地区：[1]长江大学动物科学学院,湖北荆州434025 [2]吉林大学畜牧兽医学院,吉林长春130062

出　　处：《长江大学学报（自然科学版）》2011年第3期233-235,1,共3页Journal of Yangtze University(Natural Science Edition)

摘　　要：为构建融合表达PBP39-P276的E.coli-分枝杆菌穿梭载体,分别以布氏杆菌(Brucella abortus)基因组DNA和PMD 18-T-p276质粒为模板,通过PCR扩增得到约1206bp的pbp39抗原基因和276bp的p276基因序列,将pbp39基因与穿梭表达质粒pMV361重组,得到重组质粒pMV361-pbp39,再将p276基因序列克隆至pMV361-pbp39中,得到重组质粒pMV361-pbp39-p276。质粒pMV361-pbp39-p276经PCR扩增鉴定证实,克隆基因pbp39和抗原基因p276正确插入载体pMV361。重组质粒pMV361-pbp39-p276可望在卡介苗(BCG)中分泌性表达布氏杆菌的免疫保护性抗原蛋白PBP39-P276,该质粒的构建成功为发展新型布氏杆菌疫苗奠定了基础。In order to construct the E.coli-Mycobacterium shuttle vector expressing Periplasmic binding protein (PBP39) and P276. pbp39 sequence and p276 gene were amplified from the genome of Brucella and plasmid of PMD 18-T- p276 by PCR respectively. pbp39 gene was cloned in E.coli-BCG shuttle-plasmid pMV361 to get pMV361-pbp39. Then a new recombinant plasmid pMV361-pbp39-p276 was constructed by inserting p276 into pMV361-pbp39. The results showed the cloned genes pbp39 sequence and p276 were correctly inserted into the vector pMV361, which was confirmed by PCR amplification of pMV361-pbp39-p276. pMV361-pbp39-p276 was expected to secretively express protective antigen pbp39-p276 of brucella in BCG(Bacille Calmette-guerin). It provides the possibility of further researches on the development of new anti-brucella vaccine.

关 键 词：布氏杆菌(Brucella abortus) pbp39 p276 穿梭质粒 

分 类 号：Q784[生物学—分子生物学]