Bechhofer D H, Figurski D H
Nucleic Acids Res. 1983 Nov 11;11(21):7453-69. doi: 10.1093/nar/11.21.7453.
From our earlier work, we know that the korA gene of broad host range plasmid RK2 is located within the 50.4'-56.4' region. By additional subcloning of this region, we have mapped korA to the segment between the HaeII site at 55.0' and the HincII site at 55.6'. The direction of korA transcription (55.6' to 55.1') was determined by two methods: (1) inactivation of korA expression signals and fusion of the structural gene to other promoters; and (2) hybridization analysis of korA-specific RNA's synthesized in vivo. We have determined the nucleotide sequence of the korA region. A potentially strong promoter overlaps the HincII site at 55.6', and there is a coding region which specifies the putative korA polypeptide. That this is the korA gene was supported by sequence analysis of Ba131-generated deletion mutants of korA. The sequence shows the korA product to be a small, basic polypeptide of 101 amino acids.
从我们早期的研究工作中,我们了解到广宿主范围质粒RK2的korA基因位于50.4' - 56.4'区域内。通过对该区域进行额外的亚克隆,我们已将korA定位到55.0'处的HaeII位点与55.6'处的HincII位点之间的片段。korA转录方向(从55.6'至55.1')通过两种方法确定:(1)使korA表达信号失活并将结构基因与其他启动子融合;(2)对体内合成的korA特异性RNA进行杂交分析。我们已确定了korA区域的核苷酸序列。一个潜在的强启动子与55.6'处的HincII位点重叠,并且存在一个编码区,该编码区指定了推定的korA多肽。通过对korA的Ba131产生的缺失突变体进行序列分析,证实这就是korA基因。该序列显示korA产物是一个由101个氨基酸组成的小的碱性多肽。