Garnant M K, Stauffer G V
Mol Gen Genet. 1984;193(1):72-5. doi: 10.1007/BF00327416.
Plasmid pserB59-1 carries the E. coli serB gene on a 5.2 kb BamHI fragment cloned into the BamHI site of plasmid pBR322. The results of genetic and biochemical experiments established that a functional serB gene is contained in the fragment. The location of the serB gene within the insert was determined by restriction endonuclease analysis of plasmids derived from pserB59-1 that carry the Tn5 element at sites that inactivate the serB gene, and by deletions of segments of the 5.2 kb insert that either inactivate or do not inactivate the serB gene. A 38,000 Mr serB+ polypeptide was detected when plasmid pserB59-1 was used as template in a minicell system, but not when the serB gene was inactivated by insertion of a Tn5 element.
质粒pserB59 - 1在一个5.2 kb的BamHI片段上携带大肠杆菌serB基因,该片段克隆到质粒pBR322的BamHI位点。遗传和生化实验结果表明,该片段中含有一个功能性的serB基因。通过对源自pserB59 - 1的质粒进行限制性内切酶分析来确定serB基因在插入片段中的位置,这些质粒在使serB基因失活的位点携带Tn5元件,同时通过缺失5.2 kb插入片段中使serB基因失活或不失活的片段来确定。当在小细胞系统中使用质粒pserB59 - 1作为模板时,检测到一条38,000 Mr的serB +多肽,但当serB基因通过插入Tn5元件而失活时则未检测到。