Murialdo H, Davidson A, Chow S, Gold M
Nucleic Acids Res. 1987 Jan 12;15(1):119-40. doi: 10.1093/nar/15.1.119.
Nu1 and A, the genes coding for bacteriophage lambda DNA terminase, rank among the most poorly translated genes expressed in E. coli. To understand the reason for this low level of translation the genes were cloned into plasmids and their expression measured. In addition, the wild type DNA sequences immediately preceding the genes were reduced and modified. It was found that the elements that control translation are contained in the 100 base pairs upstream from the initiation codon. Interchanging these upstream sequences with those of an efficiently translated gene dramatically increased the translation of terminase subunits. It seems unlikely that the rare codons present in the genes, and any feature of their mRNA secondary structure play a role in the control of their translation. The elimination of cos from plasmids containing Nu1 and A also resulted in an increase in terminase production. This result suggests a role for cos in the control of late gene expression. The terminase subunit overproducer strains are potentially very useful for the design of improved DNA packaging and cosmid mapping techniques.
编码噬菌体λDNA末端酶的基因Nu1和A,是在大肠杆菌中表达的翻译效率最低的基因之一。为了弄清楚翻译水平低的原因,将这些基因克隆到质粒中并检测其表达情况。此外,对这些基因起始密码子之前紧邻的野生型DNA序列进行了缩短和修饰。结果发现,控制翻译的元件存在于起始密码子上游的100个碱基对中。将这些上游序列与高效翻译基因的上游序列互换,可显著提高末端酶亚基的翻译水平。这些基因中存在的稀有密码子及其mRNA二级结构的任何特征,似乎都未在其翻译控制中发挥作用。从含有Nu1和A的质粒中去除粘性末端位点(cos),也会导致末端酶产量增加。这一结果表明cos在晚期基因表达控制中发挥作用。末端酶亚基过量生产菌株对于改进DNA包装和粘粒作图技术的设计可能非常有用。