Nikolenko G N, Kravchenko V V
Mol Gen Mikrobiol Virusol. 1996 Jan-Mar(1):28-32.
A number of p50FNL plasmids with a polycistronic operon IX-VIII-IFN-LacZ have been constructed. These plasmids provide the synthesis of polycistronic mRNA translational terminator of gene IFN situated in the coding region of lacZ gene at 41 nucleotide downstreams from its initiation codon. The coding region of IFN gene has been altered by insertions, substitutions, and deletions which provided the formation in the region of local secondary structures differing only by the loop size. The effect of this characteristic of mRNA secondary structure on lacZ translation efficiency has been assessed by measuring the levels of beta-Gal synthesis in E. coli harboring the corresponding plasmids. The level of lacZ expression increased with increase of the loop size. These data confirm that the rate of formation of mRNA secondary structure in the region of translation initiation has a crucial effect on the translation efficacy.
已经构建了许多带有多顺反子操纵子IX - VIII - IFN - LacZ的p50FNL质粒。这些质粒可在位于lacZ基因编码区起始密码子下游41个核苷酸处合成多顺反子mRNA翻译终止子。IFN基因的编码区通过插入、替换和缺失进行了改变，这些改变使得在局部二级结构区域形成了仅环大小不同的结构。通过测量携带相应质粒的大肠杆菌中β - 半乳糖苷酶的合成水平，评估了mRNA二级结构的这一特性对lacZ翻译效率的影响。lacZ表达水平随着环大小的增加而升高。这些数据证实，翻译起始区域mRNA二级结构的形成速率对翻译效率具有关键影响。
