Lewis J P, Macrina F L
Institute of Oral and Craniofacial Molecular Biology, Virginia Commonwealth University, Richmond, Virginia 23298-0566, USA.
Infect Immun. 1999 May;67(5):2619-23. doi: 10.1128/IAI.67.5.2619-2623.1999.
We have mapped a group of virulence genes of Porphyromonas gingivalis to a single large fragment of the genome. These genes (rgpA, kgp, and hagA) all contain a consensus repeat sequence (HArep). rgpA and kgp encode cysteine proteases with Arg-X and Lys-X specificity, respectively, and hagA encodes a hemagglutinin. Genomic DNA fragments separated by pulse-field gel electrophoresis were blotted and probed in order to localize the genes to a 0.25-Mb NheI fragment of the P. gingivalis W83 genome. Further hybridization analyses with single- and double-restriction digestion allowed us to generate a physical map of the fragment and determine the precise locations of the protease and hemagglutinin genes. In addition, we found an insertion-like sequence, IS195, near the ends of the 0. 25-Mb NheI fragment. A similarly sized fragment carrying HArep sequences was also demonstrated in the P. gingivalis W12 and W50 genomes.
我们已将牙龈卟啉单胞菌的一组毒力基因定位到基因组的一个单一大片段上。这些基因(rgpA、kgp和hagA)均含有一个共有重复序列(HArep)。rgpA和kgp分别编码具有Arg-X和Lys-X特异性的半胱氨酸蛋白酶,而hagA编码一种血凝素。通过脉冲场凝胶电泳分离的基因组DNA片段经印迹和杂交,以便将这些基因定位到牙龈卟啉单胞菌W83基因组的一个0.25-Mb NheI片段上。进一步用单酶切和双酶切进行杂交分析,使我们能够构建该片段的物理图谱,并确定蛋白酶和血凝素基因的精确位置。此外,我们在0.25-Mb NheI片段末端附近发现了一个类似插入序列的IS195。在牙龈卟啉单胞菌W12和W50基因组中也证实了一个携带HArep序列的大小相似的片段。