ERIC-PCR及全自动核糖体分型法鉴定不同来源的解朊金黄杆菌

目的:为了建立一种高效分离筛选高产蛋白质谷氨酰胺酶的解朊金黄杆菌的筛选方法,采用肠道基因间重复序列扩增(ERIC-PCR)和全自动核糖体分型方法对来自不同环境的33株解朊金黄杆菌(Chryseobacterium proteolyticum)进行分子分型研究。方法:采用ERIC国际通用引物对33株解朊金黄杆菌进行PCR扩增和琼脂糖凝胶电泳检测,使用NTsys软件对电泳图进行聚类分析。同时,采用全自动核糖体分型方法对33株解朊金黄杆菌进行核糖体分型,采用Bionumeric软件对菌株进行聚类分析。结果:两种方法均可得到清晰的指纹图谱,可对33株解朊金黄杆菌进行分子分型。ERIC-PCR可扩增出3~9个100~10000 bp之间的条带,将菌株分为2个族群。全自动核糖体分型方法可将菌株分为2个亚型,每种亚型间菌株仍有细微差异。结论:同种菌株之间同源性达到99%以上,但仍在遗传进化关系上存在差异。来自同一地区的分离株有聚类成一类的趋势,且与菌株的产酶能力存在密切关联。研究结果表明聚类于族群Ⅱ的菌株基本都来自于河南,且该类群菌株的产酶能力明显高于族群Ⅰ。

Identification of Chryseobacterium proteolyticum from Different Regions Using ERIC-PCR and Automated Ribotyping

Objective:In order to establish a method for efficiently isolation and screening Chryseobacterium proteolyticum strains with high ability of producing protein glutaminase,enterobacterial repetitive intergenic consensus(ERIC-PCR)and automated ribotyping(RiboPrinter)are used to study the typing of 33 bacteria strains isolated from different regions in molecular level.Methods:ERIC universal primers were used to amplify the repeated sequences that should be detected by agarose gel electrophoresis.The NTsys software was used for cluster analysis of electrophor etogram.At the same time,16S rDNA of 33 strains were classified by automatic ribosome typing method and were clustered by Bionumeric software.Results:The fingerprints of 33 strains were genotyped by both methods.ERIC-PCR could amplify 3~9 bands ranging from 100 bp to 10000 bp,and the strains were divided into two groups.Automatic ribosome typing method also can be divided into two subtypes of strains,but there still exist minor differences among each subtype of strains.Conclusion:Although the homdogy among strains belong to the same genus reached over 99%,there are still some differences on genetic evolution.At the same time,the isolates from the same region tend to cluster into one group,and a close association was exist between the enzyme production ability and typing of the strains.The results showed that all the isolates clustered in groupⅡcame from Henan Province,and the enzyme producing ability of strains in this group was significantly higher than groupⅠ.