摘要
根据国外发表的 4种来源不同的人尿道致病性大肠杆菌 pap A基因序列 ,在其保守区设计了带有 Sac /Bam H 酶切位点及保护碱基的 1对引物 ,应用 PCR从 1株带有 MRHA特性的鸡大肠杆菌染色体上扩增到 1个阳性产物 ,经酶切、连接、转化和筛选 ,得到 PCR产物的阳性克隆。通过对 PCR产物核酸序列测定 ,确证所扩增和克隆的 DNA片段为 pap
According to four published papA nucleotide sequences from different uropathogenic Escherichia coli of human,a pair of primers were designed and synthesized which were modified with Sac Ⅰ of Bam HⅠ enzyme sites.An 541 bp DNA fragment was generated by polymerse chain reaction(PCR) from an avian Escherichia coli strain expressing MRHA.The nucleotide sequence was analysed.The 541 bp DNA fragment was proved to be papA gene,which has been successfully cloned into pUC19 plasmid vector by ligation,transformation and screen by PCR and enzyme digest.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2000年第5期455-458,共4页
Chinese Journal of Veterinary Science
基金
湖北省教育委员会科学基金资助项目
