Dempsey J A, Litaker W, Madhure A, Snodgrass T L, Cannon J G
Department of Microbiology and Immunology, University of North Carolina School of Medicine, Chapel Hill 27599-7290.
J Bacteriol. 1991 Sep;173(17):5476-86. doi: 10.1128/jb.173.17.5476-5486.1991.
A physical map of the chromosome of Neisseria gonorrhoeae FA1090 has been constructed. Digestion of strain FA1090 DNA with NheI, SpeI, BglII, or PacI resulted in a limited number of fragments that were resolved by contour-clamped homogeneous electric field electrophoresis. The estimated genome size was 2,219 kb. To construct the map, probes corresponding to single-copy chromosomal sequences were used in Southern blots of digested DNA separated on pulsed-field gels, to determine how the fragments from different digests overlapped. Some of the probes represented identified gonococcal genes, whereas others were anonymous cloned fragments of strain FA1090 DNA. By using this approach, a macrorestriction map of the strain FA1090 chromosome was assembled, and the locations of various genetic markers on the map were determined. Once the map was completed, the repeated gene families encoding Opa and pilin proteins were mapped. The 11 opa loci of strain FA1090 were distributed over approximately 60% of the chromosome. The pil loci were more clustered and were located in two regions separated by approximately one-fourth of the chromosome.
已构建了淋病奈瑟菌FA1090染色体的物理图谱。用NheI、SpeI、BglII或PacI消化菌株FA1090的DNA，产生了数量有限的片段，这些片段通过轮廓夹恒定电场电泳得以分离。估计基因组大小为2219 kb。为构建该图谱，在脉冲场凝胶上分离的消化DNA的Southern印迹中使用了与单拷贝染色体序列对应的探针，以确定不同消化产生的片段如何重叠。一些探针代表已鉴定的淋球菌基因，而其他探针是菌株FA1090 DNA的无名克隆片段。通过使用这种方法，组装了菌株FA1090染色体的宏观限制性图谱，并确定了图谱上各种遗传标记的位置。图谱完成后，对编码Opa和菌毛蛋白的重复基因家族进行了定位。菌株FA1090的11个opa位点分布在染色体的约60%区域。菌毛位点更为聚集，位于被染色体约四分之一区域隔开的两个区域。
