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R质粒538-1 DNA在大肠杆菌中的体内转录与翻译

In vivo transcription and translation of R-plasmid 538-1 DNA in Escherichia coli.

作者信息

Alton N K, Vapnek D

出版信息

Contrib Microbiol Immunol. 1979;6:122-36.

PMID:231496
Abstract

In vivo transcription and translation of R-plasmid 538-1 in E. coli was analyzed. Transcription of individual restriction fragments was determined qualitatively by utilizing the techniques developed by Southern (27), and quantitatively by carrying out DNA-RNA filter hybridization. The most active region of R-plasmid transcription in strains repressed for conjugal transfer was found to occur in the region of the R-plasmid carrying the antibiotic resistance genes. In plasmids derepressed for conjugal transfer, a high level of transcription from the transfer gene region was also observed. When strains carrying R538-1drd were induced with Hg++ a high level of transcription was observed from the region of the R-plasmid carrying the genes for Hgr. Hybrid ColE1 plasmids carrying restriction fragments from the antibiotic resistance region of R538-1 were segregated into minicells. Labeling of the minicells with 35S-methionine allowed identification of the proteins coded by the fragments. A limited number of proteins were detected, and several of these have been correlated with the antibiotic resistance genes carried by R538-1.

摘要

对R质粒538 - 1在大肠杆菌中的体内转录和翻译进行了分析。利用Southern(27)开发的技术对各个限制性片段的转录进行定性测定,并通过进行DNA - RNA滤膜杂交进行定量测定。在抑制接合转移的菌株中,R质粒转录最活跃的区域发现在携带抗生素抗性基因的R质粒区域。在解除接合转移抑制的质粒中,也观察到来自转移基因区域的高水平转录。当用Hg++诱导携带R538 - 1drd的菌株时,在携带Hgr基因的R质粒区域观察到高水平转录。携带来自R538 - 1抗生素抗性区域限制性片段的杂种ColE1质粒被分离到微小细胞中。用35S - 甲硫氨酸标记微小细胞可以鉴定由这些片段编码的蛋白质。检测到数量有限的蛋白质,其中几种已与R538 - 1携带的抗生素抗性基因相关联。

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