Mashko S V, Podkovyrov S M, Trukhan M E, Lebedeva M I, Lapidus A L
Mol Biol (Mosk). 1987 Jan-Feb;21(1):73-86.
The trpOP, lacUV5, tacOP, PR and PL-promotors were cloned in the previously obtained pML4 vector plasmid. The expression of structural gene cat was studied by the chloramphenicolacetyltransferase determination in cell extracts. The level of protein synthesis by appropriate recombinant plasmids was analysed in vivo and in vitro. It was shown that the efficiency of the gene expression is determined by both the "strength" of the promotors and mRNA translation specificity. The obtained collection of the plasmids might be used for the determination of the promotor strength by the hybridization of pulse-labeled mRNA with DNA and an effective expression of the genes by means of "hybrid protein gene" and "hybrid operon" constructions.
将trpOP、lacUV5、tacOP、PR和PL启动子克隆到先前获得的pML4载体质粒中。通过测定细胞提取物中的氯霉素乙酰转移酶来研究结构基因cat的表达。在体内和体外分析了合适的重组质粒的蛋白质合成水平。结果表明,基因表达效率由启动子的“强度”和mRNA翻译特异性共同决定。所获得的质粒文库可用于通过脉冲标记的mRNA与DNA杂交来测定启动子强度,以及通过“杂交蛋白基因”和“杂交操纵子”构建来实现基因的有效表达。