Rozinov M N, Shumakov Iu L, Daĭn A A, Gol'tsmaĭer T A, Gershanovich V N
Zh Mikrobiol Epidemiol Immunobiol. 1988 Apr(4):90-3.
The 4.7 Kb EcoRI-fragment of phase I B. pertussis 475 (serovar 1.2.3) chromosome DNA carrying the pertussis toxin (PT) operon was cloned on vector plasmid pUC19 in Escherichia coli. Three fragments (1.14 Kb KpnI-PstI, 1.27 Kb PstI-PstI, and 0.96 Kb PstI-PstI) were obtained from the resulting hybrid plasmid, coded pRH119, by electrophoretic techniques and used as a combined molecular probe for analysis of the EcoRI-digested and PstI-digested chromosomal DNA of B. pertussis strain 475 in phase I, B. pertussis in phase IV, B. parapertussis strains 504 and 17903, B. bronchiseptica strain 214, and B. parapertussis strain 17903 (a convertant obtained by means of B. pertussis phage 134), as well as B. pertussis phage 134. Southern blot hybridization under the conditions of 100% DNA-DNA homology showed the presence of DNA sequences characteristic of the PT operon in all cases except the DNA of phage 134; moreover, the use of the above-mentioned probe made it possible to hybridize all EcoRI-fragments of chromosomal DNA, having the same molecular size (4.7 Kb). Consequently, the PT genes in the above Bordetella species were mapped in identical loci.(ABSTRACT TRUNCATED AT 250 WORDS)
携带百日咳毒素(PT)操纵子的I相百日咳博德特氏菌475(血清型1.2.3)染色体DNA的4.7 Kb EcoRI片段被克隆到大肠杆菌的载体质粒pUC19上。通过电泳技术从所得的杂交质粒(编码为pRH119)中获得了三个片段(1.14 Kb KpnI - PstI、1.27 Kb PstI - PstI和0.96 Kb PstI - PstI),并将其用作组合分子探针,用于分析I相百日咳博德特氏菌菌株475、IV相百日咳博德特氏菌、副百日咳博德特氏菌菌株504和17903、支气管败血博德特氏菌菌株214以及副百日咳博德特氏菌菌株17903(通过百日咳博德特氏菌噬菌体134获得的转化体)的EcoRI酶切和PstI酶切染色体DNA,以及百日咳博德特氏菌噬菌体134。在100% DNA - DNA同源性条件下的Southern印迹杂交显示,除了噬菌体134的DNA外,在所有情况下都存在PT操纵子的特征性DNA序列;此外,使用上述探针能够与染色体DNA的所有具有相同分子大小(4.7 Kb)的EcoRI片段杂交。因此,上述博德特氏菌属中的PT基因被定位在相同的位点。(摘要截短于250字)