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莱姆病病原体伯氏疏螺旋体的线性染色体物理图谱和遗传图谱。

Linear chromosomal physical and genetic map of Borrelia burgdorferi, the Lyme disease agent.

作者信息

Casjens S, Huang W M

机构信息

Department of Cellular, Viral and Molecular Biology, University of Utah Medical Center, Salt Lake City 84132.

出版信息

Mol Microbiol. 1993 May;8(5):967-80. doi: 10.1111/j.1365-2958.1993.tb01641.x.

Abstract

A physical map of the 952 kbp chromosome of Borrelia burgdorferi Sh-2-82 has been constructed. Eighty-three intervals on the chromosome, defined by the cleavage sites of 15 restriction enzymes, are delineated. The intervals vary in size from 96 kbp to a few hundred bp, with an average size of 11.5 kbp. A striking feature of the map is its linearity; no other bacterial groups are known to have linear chromosomes. The two ends of the chromosome do not hybridize with one another, indicating that there are no large common terminal regions. The chromosome of this strain was found to be stable in culture; passage 6, 165 and 320 cultures have identical chromosomal restriction maps. We have positioned all previously known Borrelia burgdorferi chromosomal genes and several newly identified ones on this map. These include the gyrA/gyrB/dnaA/dnaN gene cluster, the rRNA gene cluster, fla, flgE, groEL (hsp60), recA, the rho/hip cluster, the dnaK (hsp70)/dnaJ/grpE cluster, the pheT/pheS cluster, and the genes which encode the potent immunogen proteins p22A, p39 and p83. Our electrophoretic analysis detects five linear and at least two circular plasmids in B. burgdorferi Sh-2-82. We have constructed a physical map of the 53 kbp linear plasmid and located the operon that encodes the two major outer surface proteins ospA and ospB on this plasmid. Because of the absence of functional genetic tools for this organism, these maps will serve as a basis for future mapping, cloning and sequencing studies of B. burgdorferi.

摘要

已经构建了伯氏疏螺旋体Sh-2-82 952千碱基对染色体的物理图谱。染色体上由15种限制酶的切割位点定义的83个区间被描绘出来。这些区间大小从96千碱基对到几百碱基对不等,平均大小为11.5千碱基对。该图谱的一个显著特征是其线性;已知没有其他细菌群体具有线性染色体。染色体的两端彼此不杂交,这表明不存在大的共同末端区域。发现该菌株的染色体在培养中是稳定的;第6代、165代和320代培养物具有相同的染色体限制图谱。我们已将所有先前已知的伯氏疏螺旋体染色体基因以及几个新鉴定的基因定位在该图谱上。这些基因包括gyrA/gyrB/dnaA/dnaN基因簇、rRNA基因簇、fla、flgE、groEL(hsp60)、recA、rho/hip簇、dnaK(hsp70)/dnaJ/grpE簇、pheT/pheS簇以及编码强免疫原蛋白p22A、p39和p83的基因。我们的电泳分析检测到伯氏疏螺旋体Sh-2-82中有5个线性质粒和至少2个环状质粒。我们构建了53千碱基对线性质粒的物理图谱,并将编码两种主要外表面蛋白ospA和ospB的操纵子定位在该质粒上。由于该生物体缺乏功能性遗传工具,这些图谱将作为未来伯氏疏螺旋体图谱绘制、克隆和测序研究的基础。

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