牙龈卟啉菌rRNA基因间隔区的序列测定

Strain idendification of Porphyromonas gingivalis by sequencing analy sis of ribosomal intergenic spacer region

目的 通过rRNA基因间隔区碱基序列测定 ,建立对牙龈卟啉菌 (Pg)菌株进行鉴定的技术。 方法 利用种特异性引物对PgrRNA基因间隔区进行套式PCR扩增 ,用 glass -milk纯化PCR扩增产物 ,对纯化的扩增产物进行碱基序列测定。结果 琼脂糖凝胶电泳结果显示 ,PgrRNA基因间隔区PCR扩增产物为0 .88kb ,经放射活性测序反应技术测得PgrRNA基因间隔区的碱基序列。结论 rRNA基因间隔区DNA扩增后进行测序 ,对Pg菌株鉴定是一种精确、重复性好的分子遗传学方法。

Objective To set up a method for stra in identification of Porphyromonas gingivalis(Pg)by sequencing analysis of ribosom al intergenic spacer region.Methods Genomic DNA isolated from laboratory strain(Pg)wa s used as a template for PCR.PCR products with ribosomal intergenic spacer regio n were purified via the geneclean protocol and sequenced with Ampli cycle TM seq uencing Kit for radioactive sequencing reactions.Results The amplified products appeared at approximatery 0.88kb.The base sequence of ribosomal intergenic spacer region had been determin ed.Conclusion Ribosomal intergenic spacer region sequencing analysis provides a strain identifier for Pg that can be easily reproduced among laboratories.