细菌array-CGH技术用于比较牙龈卟啉单胞菌不同菌株基因组差异的研究  

Utilization of Bacterial Array-CGH to Compare the Differences of Genomic in Different Strains of Porphyromonas Gingivalis

作　　者：李心悦[1] 张亮[2] 吕莉娟[3] 马健[2] 周伟平[2] 黄淑君[2] 周香城 贺俊成 叶宁[4] 

机构地区：[1]广州医科大学附属广东省妇儿医院口腔科,广东广州511442 [2]广东省妇幼保健院转化医学中心,广东广州511442 [3]广东省妇幼保健院产科,广东广州511442 [4]广东省妇幼保健院口腔科,广东广州511442

出　　处：《口腔医学研究》2016年第5期506-509,共4页Journal of Oral Science Research

摘　　要：目的:构建牙龈卟啉单胞菌(简称P.gingivalis)全基因组规模的微阵列比较基因组杂交(简称arrayCGH)芯片平台,分析不同菌株间基因组的差异,为后续检测临床分离株的毒力基因,进一步阐述牙周炎发病机制提供依据。方法:综合已经全基因组测序的12株P.gingivalis菌的序列信息,设计探针序列,定制芯片。提取P.gingivalis高毒力株W83和低毒力株ATCC 33277的基因组DNA,利用array-CGH检测其全基因组的差异DNA片段,并运用PCR验证结果的准确性。结果:Array-CGH结果显示两组菌株间存在几个连续片段的拷贝数不同。P.gingivalis W83的部分特异片段参与编码毒力致病因子。在P.gingivalis ATCC 33277的特有片段中,部分编码蛋白可增强细菌表明粘附力,使其具有高粘附力。从中挑选12个基因进行PCR差异性验证,证实与芯片结果一致。结论:用array-CGH芯片的方法来分析全基因组拷贝数的变化具有分辨率高,能精确定位异常片段的特性。本文成功构建了array-CGH技术检测P.gingivalis不同毒力株基因差异的平台,为后续比较临床分离株的差异DNA片段,筛查毒力基因,深入阐述牙周炎的发病机制奠定基础。Objective：To identify the genomic differences among different strains of Porphyromonas gingivalis（P.gingivalis）to provide a basis for further exploration of periodontitis.Methods：According to the whole-genome sequencing of 12 P.gingivalis strains,the probes and the whole-genomic chip were designed.After extracting the genomic DNA of highly toxic strain P.gingivalis W83 and minimally toxic strain P.gingivalis ATCC 33277,arrayCGH was used to compare the differences and PCR was employed to verify the array results and accuracy.Results：This global comparative analysis showed the difference of continuous fragments between W83 and ATCC 33277,and the unique genes in W83 might contribute to the virulence and associate with the invasion ability of P.gingivalis.Verification with PCR confirmed the consistency with array-CGH.Conclusion：Array-CGH is an effective method to compare the genomic differences among clinical strains of periodontitis.

关 键 词：牙龈卟啉单胞菌 微阵列比较基因组杂交 基因组差异 

分 类 号：R780.2[医药卫生—口腔医学]