两种Actinobacteria鉴定方法的比较

Actinobacteria classis nov.一般包括具有超过50%G C的DNA碱基组成的微生物。其中的一些菌种在生物技术和医药方面具有重要的意义。为了分离能产生有用物质的新型微生物，我们扩大了分离对象，从以前的Streptomyes和Streptomyces以外的放线菌扩大到所有的Actinobacteria。本论文试验中的Acti-nobacteria将特指那些具有普通细菌形态的高G C含量革兰氏阳性细菌。为了能够将Actinobacteria和勘察革兰氏阳性低G C含量细菌加以区分，我们建立了两种鉴定方法。一种是荧光原位杂交（FISH）方法，它具有准确和直观的优点。另外一种是PCR方法，因在Actinobacteria 23S rRNA基因的domainⅡ（螺旋区54a）有1个大约100个碱基的稳定插入片段，通过体外PCR扩增23SrDNA片段，并用琼脂糖电泳分析，可以简便地鉴定Actinobacteria。

Compare of two methods for identifying Actinobacteria

The lineage Actinobacteria class nov. comprises bacteria with a DNA base composition which generally is above 50% G C (with a few exceptions) and are shown gram positive staining. Some species of Actinobacteria have biotechnological or medical importance. Since the discovery of Streptomycin, a large number of antibiotics have been isolated from the cultures of a wide variety of Streptomycetes and rare Actinomycetes . For increasing the chance of discovering new useful compounds from microbial, we expanded the scope of screening targets from the members of Actinomycetes to Actinobacteria . Two methods were set up, such as FISH and PCR, to identify Actinobacteria . In order to confirm the accuracy of FISH and PCR result, the DNA G C contents of those strains were determined. It was showed that the results of G C content determination and both of two methods were identical. Comparing these two methods, FISH identification method was effective and reliable, while PCR method was more simple to carry out.