LeBlanc D J, Inamine J M, Lee L N
Antimicrob Agents Chemother. 1986 Apr;29(4):549-55. doi: 10.1128/AAC.29.4.549.
The Emr, Kmr, and Smr determinants of the Streptococcus faecalis R plasmid pJH1 were cloned in Streptococcus sanguis with a streptococcal plasmid vector, pVA380-1. Each cloned determinant was used as a probe in hybridization reactions with dot blots containing plasmid-enriched DNA from 91 group D streptococcal isolates resistant to erythromycin, kanamycin, and streptomycin; the isolates were obtained from animal and human sources in a variety of geographical locations. Nearly 70% of the strains contained DNA that hybridized to each of the three resistance determinants from pJH1. Five plasmids mediating resistance to erythromycin, kanamycin, and streptomycin were examined in more detail. These plasmids varied in size between 26 and 105 kilobase pairs (kbp) and exhibited very different EcoRI restriction patterns. However, each plasmid contained the resistance determinants on a single 13- to 20-kbp EcoRI fragment. Southern blot hybridizations and additional restriction endonuclease digests revealed extensive DNA sequence homology and virtually indistinguishable restriction endonuclease maps within a 9- to 11-kbp region of each plasmid which included the resistance determinants.
粪肠球菌R质粒pJH1的Emr、Kmr和Smr决定簇用链球菌质粒载体pVA380 - 1克隆到血链球菌中。每个克隆的决定簇在与包含来自91株对红霉素、卡那霉素和链霉素耐药的D群链球菌分离株的富含质粒的DNA的斑点杂交反应中用作探针;这些分离株来自不同地理位置的动物和人类。近70%的菌株含有与pJH1的三种耐药决定簇中的每一种杂交的DNA。对5种介导对红霉素、卡那霉素和链霉素耐药的质粒进行了更详细的研究。这些质粒大小在26至105千碱基对(kbp)之间变化,并且表现出非常不同的EcoRI限制性图谱。然而,每个质粒在一个单一的13至20 kbp的EcoRI片段上含有耐药决定簇。Southern印迹杂交和额外的限制性内切酶消化揭示了每个质粒的一个9至11 kbp区域内广泛的DNA序列同源性和几乎无法区分的限制性内切酶图谱,该区域包括耐药决定簇。