Hobbs M, Dalrymple B, Delaney S F, Mattick J S
Department of Biotechnology, University of New South Wales, Kensington, Australia.
Gene. 1988;62(2):219-27. doi: 10.1016/0378-1119(88)90560-4.
Gene fimA encoding the structural subunit of the fimbriae of Pseudomonas aeruginosa PAK is located in the centre of a 1.2-kb HindIII genomic DNA fragment [see also Sastry et al., J. Bacteriol. 164 (1985) 571-577], which in turn is located within a 6.2-kb EcoRI fragment. Immediately downstream from fimA is a putative threonine tRNA gene [Dalrymple and Mattick, Biochem. Int. 13 (1986) 547-553]. Northern blotting experiments showed that fimA is transcribed to an mRNA of approx. 650 nucleotides, which also includes the threonine tRNA sequence but no other protein-coding region. There was no indication that this mRNA is processed to release the tRNA sequence. However, the tRNA did appear to be expressed independently from its own promoter in the region 3' to fimA. When these sequences were introduced into Pseudomonas putida, we found that the level of expression of fimA from the cloned 6.2-kb EcoRI fragment was approx. 30-fold greater than that from the smaller HindIII fragment, whereas that of the specific tRNA species was unaltered. The size of the fimA transcript was also unaltered. These results provide evidence that the fimA gene is subject to specific transcriptional activation in vivo and that this activation involves sequences flanking the 1.2-kb HindIII fragment.
编码铜绿假单胞菌PAK菌毛结构亚基的基因fimA位于一个1.2kb的HindIII基因组DNA片段的中心位置[另见Sastry等人,《细菌学杂志》164 (1985) 571 - 577],而该片段又位于一个6.2kb的EcoRI片段内。fimA紧邻下游是一个假定的苏氨酸tRNA基因[Dalrymple和Mattick,《生物化学国际》13 (1986) 547 - 553]。Northern印迹实验表明,fimA转录成一个约650个核苷酸的mRNA,其中也包含苏氨酸tRNA序列,但没有其他蛋白质编码区。没有迹象表明该mRNA会被加工以释放tRNA序列。然而,tRNA似乎确实在fimA 3'区域从其自身的启动子独立表达。当将这些序列导入恶臭假单胞菌时,我们发现从克隆的6.2kb EcoRI片段中fimA的表达水平比从小的HindIII片段中高约30倍,而特定tRNA种类的表达水平未改变。fimA转录本的大小也未改变。这些结果提供了证据,表明fimA基因在体内受到特定的转录激活,并且这种激活涉及1.2kb HindIII片段侧翼的序列。