RESTRICTION MAP OF ANTIBIOTIC-RESISTANCE PLASMID R1DRD-19 AND ITS DERIVATIVES PKN102 (R1DRD-19B2) AND R1DRD-16 FOR ENZYMES BAMHI, HINDIII, ECORI AND SALI
RESTRICTION MAP OF ANTIBIOTIC-RESISTANCE PLASMID R1DRD-19 AND ITS DERIVATIVES PKN102 (R1DRD-19B2) AND R1DRD-16 FOR ENZYMES BAMHI, HINDIII, ECORI AND SALI
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DOI：
10.1007/bf00266905
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发表时间：
1978-01-01
期刊：
影响因子：
--
通讯作者：
GOEBEL, W
中科院分区：
文献类型：
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作者：
BLOHM, D;GOEBEL, W
使用限制性内切酶 BamHI、HindIII、EcoRI 和 SalI 对介导对氨苄西林 (Ap)、氯霉素 (Cm)、卡那霉素 (Km)、链霉素 (Sm) 和磺酰胺 (Su) 抗生素耐药性的接合 R 质粒 R1drd-19 [来自大肠杆菌] 进行定位。 BamHI 生成 5 个片段 (A-E)，MW 介于 46 × 之间。 106 道尔顿（主要代表 RTF [电阻传递因子]）和 0.25 倍。 106 道尔顿，HindIII 8 (A-H) 42 倍之间。 106道尔顿（代表RTF的主要部分）和0.1.times。 106 道尔顿。 EcoRI 可识别 17 个位点并产生分子量在 11.7 至 0.1 倍之间的片段 (A-Q)。 106 道尔顿。 SalI产生7个16.5至2.0倍的片段(A-G)。 106 道尔顿。通过用 1 种限制酶部分消化 R1drd-19、用 2 或 3 种酶对 DNA 进行双重和三重消化（分离和不分离）获得的片段以及来自制备凝胶的各个条带构建物理图谱。 R1drd-19的几个突变体的限制性酶切模式也与其进行了比较。所研究的R1衍生物是通过扩展缺失产生的，即失去Km抗性的拷贝突变体pKN102、失去Km以外的所有抗性的R1drd-16和R1drd-16的Kms衍生物，其代表纯RTF。 R1drd-19 的图谱与 R100 和 R6-5 的图谱明显不同。其分子量为 62.5 Md [兆道尔顿]。 BamHI 的环状片段顺序为：A-C-B-D-E，HindIII 为：A-D-C-B-F-H-E-G，EcoRI 为：A-C-K-B-F-J-O-D-H-L-G-P-Q-N-I-E-M-，SalI 为 A-B-C-D-G-F-E。
The conjugative R plasmid R1drd-19 [from Escherichia coli], mediating antibiotic resistance to ampicillin (Ap), chloramphenicol (Cm), kanamycin (Km), streptomycin (Sm) and sulfonamides (Su), was mapped using the restriction endonucleases BamHI, HindIII, EcoRI and SalI. BamHI generates 5 fragments (A-E) with MW between 46 .times. 106 dalton (representing mainly the RTF [resistance transfer factor]) and 0.25 .times. 106 dalton, and HindIII 8 (A-H) between 42 .times. 106 dalton (representing the main part of the RTF) and 0.1 .times. 106 dalton. EcoRI recognises 17 sites and produces fragments (A-Q) with MW between 11.7 and 0.1 .times. 106 dalton. SalI yields 7 fragments (A-G) of 16.5 to 2.0 .times. 106 dalton. A physical map was constructed from fragments obtained by partial digestion of R1drd-19 with 1 restriction enzyme, by double and triple digestion of the DNA with 2 or 3 enzymes with and without isolation and individual bands from preparative gels. The restriction patterns of several mutants of R1drd-19 were also compared with it. The derivatives of R1 investigated are generated by extended deletions, namely the copy mutant pKN102 which lost the Km resistance, R1drd-16, which lost all resistances other than Km and the Kms derivative of R1drd-16, which represents the pure RTF. The map of R1drd-19 is remarkably different from those of R100 and R6-5. Its MW was 62.5 Md [megadalton]. The circular fragment order for BamHI is: A-C-B-D-E, for HindIII: A-D-C-B-F-H-E-G, for EcoRI: A-C-K-B-F-J-O-D-H-L-G-P-Q-N-I-E-M- and for SalI A-B-C-D-G-F-E.