摘要
根据已成功扩增的大肠杆菌K88及K99菌毛蛋白结构基因PCR反应 ,进行PCR反应体系的优化。将 2种菌毛蛋白结构基因的扩增引物混合进行PCR ,同时检测K88及K99菌毛蛋白结构基因。经优化试验 ,筛选出 5 0 μLPCR反应体系中各成分的最适用量为 :Mg2 + 浓度 2mmol L ,Taq酶 5U ,dNTPs 2 0 0mmol L ,每条引物 5 0pmol,模板量 1 0 0CFU ,并研制成功用于临床检测的PCR诊断试剂盒。
On the basis of the technique of PCR that had amplified successfully Escherichia coli K88 and K99 pili protein structure gene, the system of PCR were optimized. The K88, K99 pili protein structure gene were detected by mixing two pairs of primers for pili protein structure genes. In the 50 μL optimal system, the template, Taq enzyme, and Mg 2+ were 100 CFU, 5 U and 2 mmol/L respectively , the amounts of each dNTP and primer were 50 mmol/L and 50 pmol respectively. At the same time, the PCR diagnosis kits which could be used in clinical practice were manufactured successfully.
出处
《湖北农学院学报》
2002年第6期501-503,510,共4页
Journal of Hubei Agricultural College
基金
湖北省教委重大攻关资助项目 (1 998第 4号 )
