Beresford T, Condon S
Department of Food Microbiology and National Food Biotechnology Centre, University College, Cork, Ireland.
FEMS Microbiol Lett. 1991 Mar 1;62(2-3):319-23. doi: 10.1016/0378-1097(91)90178-d.
A cosmid gene library of the genome of Lactococcus lactis subsp. lactis 712 was probed for the presence of 16S rRNA genes, using 32P 5' end-labelled 16S rRNA fragments. Cosmid DNA from positive clones responsible for hybridisation was subcloned into a high copy number vector and a restriction map was constructed. The location of the 16S, 23S and 5S rRNA genes was determined on this map. Transcriptional promoter activity was identified upstream of the 5' end of the 16S rRNA gene. By probing L. lactis 712 chromosomal DNA cut with a range of restriction endonucleases, with a conserved oligonucleotide to the 5' end of the 16S rRNA gene, 6 copies of rRNA genes were identified.
使用32P 5'端标记的16S rRNA片段，对乳酸乳球菌乳酸亚种712基因组的黏粒基因文库进行16S rRNA基因检测。将负责杂交的阳性克隆的黏粒DNA亚克隆到高拷贝数载体中，并构建限制酶图谱。在该图谱上确定了16S、23S和5S rRNA基因的位置。在16S rRNA基因5'端上游鉴定到转录启动子活性。通过用一系列限制性内切酶切割乳酸乳球菌712染色体DNA，并用与16S rRNA基因5'端保守的寡核苷酸进行杂交，鉴定出6个rRNA基因拷贝。
