Comstruction of a physical and gene map of periodontpathic bacterial genome

牙周病细菌基因组物理和基因图谱的构建

基本信息

批准号：
05454493
负责人：
UMEMOTO Toshio
金额：
$ 3.65万
依托单位：
Kanagawa Dental College
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1995
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05454493/
关键词：
Periodontpathic bacteria P.gingivalis Physical map NotI fragment Genome size Gene map Virulent gene Pulsed-field gel electrophoresis 歯周病原細菌 Porphyromonas gingivalis 染色体DNA 制限酵素地図遺伝子地図

项目摘要

Genetic approaches have been recently introduced to analyze virulence factors of Porphyromonas gingivalis. Several genes potentially involved in their pathogenicity related with periodontal disease. However, little is known of the over all genetic organization of this bacterium. Recently, pulsed-field gel electrophoresis was developed to separate large DNA fragment. Physical maps of several bacterial genome have been constructed by using pulsed-field gel electrophoresis. In this study, a physical and gene map of genome of P.gingivalis ATCC 33277 constructed by Southern blot analysis of NotI restriction fragments separated by pulsed-field gel electrophoresis. We firstly examined polymorphism of the patterns of the NotI-digested chromosomal DNA fragments of the strains ATCC 33277,381, ATCC 53977, W50, and W83. All these strains showed completely different patterns, though only those of W50 and W83 were very similar.Then we tried to construct a NotI map of the chromosomal DNA of P.gingivalis ATCC 33277. The restriction enzyme NotI produced at least 10 fragments, which were ranging between kilobases of 30 and 690. These sizes were determined by pulsed-field gel electrophoresis. Thus, the total size of the chromosome was estimated to be about 2600 kb. Subsequently, Southern hybridization analysis of the NotI-digested chromosome was performed with some DNA probes, such as the fimbrillin gene (fimA), the superoxide dismutase gene (sod) and one of the genes of trypsin-like protease. The probe for fimA gene hybridized to the largest fragment and that for sod to the fifth. The probe for the trypsin gene hybridized to the sixth fragment from the top. These results suggested that there was no close linkage among these three genes examined.

最近引入了遗传方法来分析牙龈毒死菌的毒力因子。几个基因可能涉及其致病性与牙周疾病有关。但是，对于该细菌的所有遗传组织，知之甚少。最近，开发了脉冲场凝胶电泳以分离大型DNA片段。通过使用脉冲场凝胶电泳构建了几个细菌基因组的物理图。在这项研究中，通过脉冲场凝胶电泳分离的Noti限制片段构建的吉利亚氏菌ATCC基因组的物理和基因图33277。我们首先检查了菌株ATCC 33277,381，ATCC 53977，W50和W83的Noti消化染色体DNA片段的多态性。所有这些菌株都显示出完全不同的模式，尽管只有W50和W83的模式非常相似。因此，染色体的总大小估计约为2600 kb。随后，用一些DNA探针进行了Noti消化染色体的南部杂交分析，例如纤维林基因（FIMA），超氧化物歧化酶基因（SOD）和胰蛋白酶样蛋白酶的基因之一。 FIMA基因的探针与最大的片段杂交，并将其引入第五。胰蛋白酶基因的探针从顶部杂交至第六片。这些结果表明，在检查的这三个基因之间没有紧密的联系。