摘要
目的构建肠出血性大肠埃希菌（EHEC）O157：H7紧密黏附素C-端免疫保护片段与肠黏膜佐剂大肠埃希菌不耐热肠毒素B亚单位（LTB）的融合基因克隆载体,为进一步研究EHEC O157：H7重组疫苗奠定了基础.方法设计引物采用PCR技术从EHEC O157：H7基因组中扩增Intimin C300的编码基因,从肠产毒性大肠埃希菌基因组中扩增LTB的编码基因,分别构建克隆载体pUC18-C300和pUC18-LTB,采用酶切位点相连技术,构建pUC19-C300-LTB克隆载体,并测序.结果从EHEC O157：H7基因组中扩增出了900 bp的目的片段,从肠产毒素大肠埃希菌基因组扩增出了275 bp的目的片段,分别插入载体pUC18和pUC19,经酶切及测序鉴定与目的序列一致.结论克隆出EHEC O157：H7的紧密黏附素C-端免疫保护片段与不耐热肠毒素B亚单位基因,并成功构建融合基因克隆载体,测序正确.
Objective To construct the clone vector of the Intimin C300-LTB from the Hemorrhage E.coli（EHEC）O157：H7 and the heat-labile enterotoxin B（LTB）.Method The primers for Intimin C300 and LTB were designed and the PCR technique was used to amplify the target genes from the genomes of EHEC O157：H7 and EHEC respectively.The clone vectors pUC18-Intimin C300,pUC18-LTB and pUC19-Intimin C300-LTB were constructed for further sequencing and identification.Results The objective fragments of 275 bp（LTB）and 900 bp（Intimin C300）from EHEC O157：H7 and EHEC were amplified by PCR.The recombinant plasmid pUC19-Intimin C300-LTB was constructed,and sequencing proved the perfect results.Conclusion The genes of LTB and Intimin C300 were cloned correctly and the pUC18-C300,pUC18-LTB and pUC19-Intimin C300-LTB clone vectors were constructed successfully.The fusion gene of Intimin C300-LTB has laid the foundation for developing new vaccine of the EHEC O157：H7.
出处
《北华大学学报（自然科学版）》
CAS
2010年第3期225-228,共4页
Journal of Beihua University（Natural Science）
基金
吉林省科技发展计划项目(200805103)
