空肠弯曲菌质粒DNA提取条件的优化

A modified extraction method of the plasmid DNA in Campylobacter Jejuni

在碱解法的基础上,优化了提取空肠弯曲菌质粒DNA的条件。细菌经37℃,5%CO2培养48h后,每个样品以长满一个φ90mm平板(约2.86×106个/mL)的细菌用量为宜;溶菌液的最适SDS浓度和pH值为2%和12.0～12.5;最适温育温度和时间为55℃、60min。在该条件下用该方法提取了219株不同畜禽来源的空肠弯曲菌质粒DNA,质粒阳性率为10.96%,最大质粒DNA108kb,最小质粒DNA1.9kb。

The conditions of extracting plasmids DNA in Campylobacterjejuni were optimized. The optimum concentration of bacteria was 2.86×10^6 cells/mL, and the optimum concentration of SDS and pH of the baeteriolysant were 2 % and 12.0-12.5 respectively. 219 strains of Campylobacter jejuni plasmid DNA from different animals were extracted under these conditions, the positive rate was 10.96 % （24/219）. The maximal plasmid DNA was 108 kb, and the minimal was 1.9 kb.