Yeh P, Sicard A M, Sinskey A J
Department of Applied Biological Sciences, Massachusetts Institute of Technology, Cambridge 02139.
Mol Gen Genet. 1988 Apr;212(1):112-9. doi: 10.1007/BF00322452.
Sequence analysis localized the lysA gene of Corynebacterium glutamicum strain AS019 within a 1.35 kb open reading frame, potentially encoding a 445 amino acid product. Immediately downstream from this gene we found a potential rho-independent transcription terminator, while the 5' flanking region (300 bp) harbors unusual topological and structural features, located in the vicinity of a potential ribosome binding site. Within this upstream region, enzymatic and genetic analyses indicated the occurrence of a promoter responsible for significant, although weak, expression of the encoded enzymatic activity. The same significant expression level was observed with a plasmid harboring an additional 0.5 kb of genomic information upstream from lysA, while its full expression apparently requires 2 kb of additional genomic information located immediately upstream from the cloned gene. The upstream sequence requirement apparently associated with the full expression of the lysA gene of C. glutamicum shows some similarity with the Escherichia coli system.
序列分析将谷氨酸棒杆菌AS019菌株的lysA基因定位在一个1.35 kb的开放阅读框内,该开放阅读框可能编码一个445个氨基酸的产物。在该基因的紧邻下游,我们发现了一个潜在的不依赖ρ因子的转录终止子,而5'侧翼区域(300 bp)具有异常的拓扑和结构特征,位于一个潜在的核糖体结合位点附近。在这个上游区域,酶学和遗传学分析表明存在一个启动子,该启动子负责编码酶活性的显著表达,尽管表达较弱。用一个在lysA上游携带额外0.5 kb基因组信息的质粒也观察到了相同的显著表达水平,而其完全表达显然需要位于克隆基因紧邻上游的2 kb额外基因组信息。谷氨酸棒杆菌lysA基因完全表达所明显需要的上游序列要求与大肠杆菌系统有一些相似性。