Blanco C, Ritzenthaler P, Kolb A
Mol Gen Genet. 1986 Jan;202(1):112-9. doi: 10.1007/BF00330526.
The uxuAB operon is composed of two genes coding for enzymes involved in hexuronate degradation. This operon is negatively controlled by the uxuR and exuR regulatory gene products. Starting from uxu hybrid plasmids, a 300 bp Sau3A restriction fragment was isolated and shown to contain the entire uxu regulatory region using in vitro gene fusions that brought lac gene expression under the control of the transcriptional and translational signals of the uxuA gene. The nucleotide sequence of this fragment was established. The start of the uxu mRNA was localized by S1 mapping experiments (the presence of a CAP binding site upstream of the promoter was shown by DNAse I footprinting and in vitro titration). The N-terminal amino acid sequence of the purified uxuA-lacZ gene product allowed the mapping of the uxuA initiation codon at 115 bp from the start of transcription.
uxuAB操纵子由两个编码参与己糖醛酸降解的酶的基因组成。该操纵子受uxuR和exuR调控基因产物的负调控。从uxu杂交质粒开始,分离出一个300 bp的Sau3A限制性片段,并通过体外基因融合证明其包含整个uxu调控区,该融合使lac基因表达受uxuA基因转录和翻译信号的控制。确定了该片段的核苷酸序列。通过S1作图实验确定了uxu mRNA的起始位置(通过DNA酶I足迹法和体外滴定法显示启动子上游存在CAP结合位点)。纯化的uxuA-lacZ基因产物的N端氨基酸序列使得能够将uxuA起始密码子定位在距转录起始点115 bp处。