摘要
根据U.Washington报导的根癌土壤农杆菌(Agribacterium tumefaciens)C58Ti质粒基因序列,设计1对引物,利用PCR的方法扩增了c58根癌土壤农杆菌Ti质粒毒性区VirD1基因。通过琼脂糖凝胶电泳,所得目的片段略小于500bp,与引物设计跨幅片段476bp相吻合。将此片段连接到T载体,经蓝白斑筛选、PCR鉴定、测序及DNA序列分析,结果表明:克隆VirD1基因编码序列与报导的序列同源性达100％,说明通过PCR方法获得的VirD1基因片段是正确的,为进一步研究该基因的表达和活性奠定了基础。
According to the Agribacterium tumefaciens' Ti plasmid gene sequences published by U. Washington, we designed a pair of primers, amplified the VirD1 protein gene situating in virus region of agribacterium tumefaciens' Ti Plasmid with the method of PCR. Though agarose electrophoresis the cloning fragment is lower a little than 500bp, which identical with the design length (476bp). Join the frament with T-vector, through blue and white spot selection, sequencing and DNA sequence analysis, the result shows:there are 100% accordance between cloning VirD1 gene coden sequence and the published gene sequence. Conclusion:Through PCR method we get the correct VirD1 protein gene sequence, this will be helpful to study the gene's translation and activity.
出处
《石河子大学学报（自然科学版）》
CAS
2004年第3期194-196,共3页
Journal of Shihezi University(Natural Science)
基金
新天石大科研基金(XS200009)
