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Detection and analysis of new functional genes on the genome of bacterio phages

噬菌体基因组新功能基因的检测与分析

基本信息

批准号：
18570001
负责人：
SADAIE Yoshito
金额：
$ 2.61万
依托单位：
Saitama University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18570001/
关键词：
Bacillus subtilis bacteriophage restriction modification system genomics

项目摘要

When the bacteriophage is infected with specific host bacteria, it become lysogeny in a host chromosome or multiplies in a host cell and finally causes lysis of a host. The prophage genes which become lysogeny been mutated afterwards, and these can bring a host a new function. Many of protein encoded by a gene on the bacteriophage DNA are novel, and we can discover useful genes of unknown function and structure. We begin this research aiming at search and analysis of such novel genes.Bacillus subtilis Maeberg is tolerance to bacteriophage SP10, but it becomes sensitive when the nonA and the nonB mutations are occurred in B. subtilis cells. We started this study to elucidate this phenomenon.I performed the sequencing of the SP10 bacteriophage genome to analyze mechanism for the infection of SP10 bacteriophage. For making librarye, SP10 genomic DNA was sheared by plural restriction enzymes and supersonic waves. These fragments were cloned into a plasmid vector and sequenced. These sequenced data were assembled and made some contig. 72 contig was provided by the present and 97, 547bp of 90% in total length (about 108kb) were determined by sequencing. High similarities with Staphylococcus phage K ORF were often observed. We tried to determine whole genome sequence now.The nonA locus is lied in SP□, B. subtilis prophage. We deleted gradually SP□ out from B. subtilis chromosome and tested their sensitivity to SP10 to determine the gene which causes nonA phenotype. As a result of this analysis, it became clear that there was plural mechanism. There are at least two mechanism concerned. One is yonO gene and non-identified genes, and another is homolog of the dNDP reductase. The function of yonO gene is unknow, but is possibly regulator of gene expression. Future analysis is still to be elucidating.

当噬菌体被特定的宿主细菌感染时，它在宿主染色体中成为溶原性的或在宿主细胞中增殖，并最终导致宿主的裂解。成为溶原性的原噬菌体基因随后发生突变，这些基因可以给宿主带来新的功能。噬菌体DNA上的基因编码的许多蛋白质都是新的，我们可以发现功能和结构未知的有用基因。本研究开始的目的是寻找和分析这些新基因，枯草芽孢杆菌Maeberg对噬菌体SP10具有耐受性，但当B发生非A和非B突变时，它变得敏感。枯草杆菌细胞为了阐明这一现象，我们进行了SP10噬菌体的基因组测序，以分析SP10噬菌体的感染机制。为了制备文库，用多种限制性内切酶和超声波剪切SP10基因组DNA。将这些片段克隆到质粒载体中并测序。将这些测序数据进行组装并制成一些重叠群。本研究共获得72个重叠群，测序结果为97，547bp，占全长（约108kb）的90%。经常观察到与葡萄球菌噬菌体K ORF高度相似。目前我们正在进行全基因组测序，非A位点位于SP□、B。枯草前噬菌体我们逐渐从B中删除SP□。subtilis染色体上分离，并检测其对SP10的敏感性，以确定引起nonA表型的基因。通过这种分析，很明显存在多元机制。至少有两种机制。一个是yonO基因和未鉴定的基因，另一个是dNDP还原酶的同源物。yonO基因的功能尚不清楚，但可能是基因表达的调节因子。未来的分析仍有待阐明。