Hespell R B, Whitehead T R
National Center for Agricultural Utilization Research, U.S. Department of Agriculture, Peoria, Illinois 61604.
Appl Environ Microbiol. 1991 Sep;57(9):2703-9. doi: 10.1128/aem.57.9.2703-2709.1991.
Anaerobic filter matings of Butyrivibrio fibrisolvens H17c, CF3, D1, or GS113, representing different DNA relatedness groups, were done with Enterococcus faecalis CG110, which contains chromosomally inserted Tn916. Tetracycline-resistant transconjugants were obtained with each mating pair at average frequencies of 4.4 x 10(-6) (per recipient) and 5.2 x 10(-6) (per donor). The transfer frequencies of Tn916 into B. fibrisolvens varied 5- to 10-fold with mating time, strain, and growth stage. By using Southern hybridization with pAM120 as the probe, Tn916 was shown to insert at one or more separate chromosomal sites for each strain of B. fibrisolvens. Retransfer of Tn916 from B. fibrisolvens H17c or CF3 to E. faecalis OG1-X or JH 2-2 or to B. fibrisolvens D1 or GS113 could not be shown. Matings of E. faecalis RH110, which contains chromosomally inserted Tn916 delta E, with B. fibrisolvens 49, H17c, D1, CF3, GS113, or VV-1 resulted in erythromycin-resistant transconjugants at average frequencies of 5.3 x 10(-7) (per recipient) and 2.5 x 10(-7) (per donor). Tn916 delta E was shown by Southern hybridization with pAM120 to insert at one or more sites in the chromosome of each strain. B. fibrisolvens H17c was anaerobically filter mated with E. faecalis JH 2-SS, which contains pAM beta 1. Erythromycin-resistant transconjugants were obtained at frequencies of 2 x 10(-5) (per recipient) and 6 x 10(-5) (per donor). The presence of pAM beta 1 in these transconjugants could not be shown by agarose gel electrophoresis of plasmid minilysates but could be shown by Southern hybridization analysis.(ABSTRACT TRUNCATED AT 250 WORDS)
用含有染色体插入Tn916的粪肠球菌CG110与代表不同DNA相关性组的溶纤维丁酸弧菌H17c、CF3、D1或GS113进行厌氧滤膜交配。每个交配组合均获得了四环素抗性的接合子,平均频率分别为4.4×10⁻⁶(每受体)和5.2×10⁻⁶(每供体)。Tn916向溶纤维丁酸弧菌的转移频率随交配时间、菌株和生长阶段的不同而变化5至10倍。通过以pAM120为探针进行Southern杂交,结果显示对于溶纤维丁酸弧菌的每个菌株,Tn916均插入到一个或多个不同的染色体位点。未显示Tn916从溶纤维丁酸弧菌H17c或CF3向粪肠球菌OG1-X或JH 2-2或向溶纤维丁酸弧菌D1或GS113的再次转移。用含有染色体插入Tn916δE的粪肠球菌RH110与溶纤维丁酸弧菌49、H17c、D1、CF3、GS113或VV-1进行交配,获得了红霉素抗性的接合子,平均频率分别为5.3×10⁻⁷(每受体)和2.5×10⁻⁷(每供体)。通过以pAM120进行Southern杂交显示,Tn916δE插入到每个菌株染色体的一个或多个位点。用含有pAMβ1的粪肠球菌JH 2-SS与溶纤维丁酸弧菌H17c进行厌氧滤膜交配。获得了频率为2×10⁻⁵(每受体)和6×10⁻⁵(每供体)的红霉素抗性接合子。通过质粒小裂解物的琼脂糖凝胶电泳未显示这些接合子中存在pAMβ1,但通过Southern杂交分析可以显示。(摘要截短于250字)