Ukhabotina L S, Belova T S, Zhukov V G, Danilenko V N
Antibiot Khimioter. 1988 Feb;33(2):87-93.
Presence of plasmid DNA was investigated in laboratory strains 2 and 4 (NRRL 2338) of S. erythreus, as well as in strains 1 and 3 of S. erythreus subjected to improvement with respect to erythromycin production. Families of plasmids close by their molecular weights were identified in S. erythreus strains 3 and 4 (NRRL 2338). A plasmid DNA fraction of S. erythreus strain 3 was studied with electron microscopy. It enabled to identify 5 plasmids: pSE11, pSE12, pSE13, pSE14 and pSE15 with length of 5.3, 12.4, 16.3, 29.6 and 86.9 kb respectively. Using of various procedures for isolation of extrachromosomal DNA did not provide its detection in S. erythreus strains 1 and 2. At least a part of the plasmids detected in S. erythreus strains 3 and 4 (NRRL 2338) was conjugative. 32R-Labeled plasmid DNA of S. erythreus strain 3 was subjected to hydridization according to Sauthern with total DNA of the 4 strains treated with restrictases BamHI, PstI and BgIII. The studies showed that the genome of S. erythreus strain 2 was not homologous with the probe while S. erythreus strain 1 contained one of the plasmids or its part in chromosome-integrated state. In strains 3 and 4 (NRRL 2338) of S. erythreus certain plasmid DNAs were present in both autonomous and chromosome-inserted states. 32P-Labeled gene of erythromycin resistance (ermE) was subjected to hybridization according to Southern with total DNA of the 4 strains and with DNA plasmid fraction of S. erythreus strain 3. The signal was positive only in hydridization of the probe with total DNA of S. erythreus strains 1, 3, and 4 (NRRL 2338).(ABSTRACT TRUNCATED AT 250 WORDS)
研究了红链霉菌实验室菌株2和4(NRRL 2338)以及在红霉素生产方面经过改良的红链霉菌菌株1和3中质粒DNA的存在情况。在红链霉菌菌株3和4(NRRL 2338)中鉴定出了分子量相近的质粒家族。对红链霉菌菌株3的质粒DNA部分进行了电子显微镜研究。鉴定出了5种质粒:pSE11、pSE12、pSE13、pSE14和pSE15,长度分别为5.3、12.4、16.3、29.6和86.9 kb。使用各种分离染色体外DNA的方法未在红链霉菌菌株1和2中检测到它。在红链霉菌菌株3和4(NRRL 2338)中检测到的至少一部分质粒是可接合的。用32R标记的红链霉菌菌株3的质粒DNA与经限制性内切酶BamHI、PstI和BgIII处理的4种菌株的总DNA进行Southern杂交。研究表明,红链霉菌菌株2的基因组与探针不同源,而红链霉菌菌株1含有一种质粒或其部分处于染色体整合状态。在红链霉菌菌株3和4(NRRL 2338)中,某些质粒DNA以自主状态和染色体插入状态存在。用32P标记的红霉素抗性基因(ermE)与4种菌株的总DNA以及红链霉菌菌株3的质粒细胞DNA部分进行Southern杂交。仅在探针与红链霉菌菌株1、3和4(NRRL 2338)的总DNA杂交时信号为阳性。(摘要截于250字)