摘要
合成O 抗原的基因是串联在一起的一个基因簇 ,提取O1 39霍乱弧菌基因组DNA ,限制性内切酶EcoRⅠ酶切 ,电泳回收 4～ 2 0kb的DNA片段 ,构建质粒基因组文库。随机筛选重组克隆 ,获得一株可与O1 39霍乱弧菌抗血清发生凝集反应的重组克隆 ,命名为大肠杆菌DH5α(pMG32 0 )。经鉴定分析重组克隆所表达的O 抗原具有良好的免疫原性及反应原性。酶切分析质粒 pMG32 0 ,推知其O 抗原基因大小约 4 6kb。这为今后O1 39霍乱疫苗的研制及O1 39霍乱弧菌O 抗原基因的结构和功能研究提供了条件。
Because O\|antigen biosynthesis genes are a tandem gene cluster.Gnomic fragments of 4～20 kilobases (kb) were obtained by digesting genomic DNA of V.cholerae O 139 with restriction endonuclease Eco RI,then plasmid gene bank was constructed.Recombinant colony, E.coli DH5α (pMG320),expressing O\|antigen of V.cholerae O 139 was detected from the bank by immunological agglutinative reaction.The futher analysis showed O\|antigen expressed by recombinant colony had both immunogenicity and reactogenicity,and the size of O\|antigen biosynthesis genes was about 4.6kb.
出处
《微生物学报》
CAS
CSCD
北大核心
2001年第1期65-69,共5页
Acta Microbiologica Sinica
基金
国家863资助项目(102-07-03-02)
