Pestka S, Daugherty B L, Jung V, Hotta K, Pestka R K
Proc Natl Acad Sci U S A. 1984 Dec;81(23):7525-8. doi: 10.1073/pnas.81.23.7525.
A plasmid was constructed to generate RNA complementary to the beta-galactosidase mRNA under control of the phage lambda PL promoter. When this anti-mRNA was produced, synthesis of beta-galactosidase was dramatically inhibited (98%). Syntheses of galactoside permease and transacetylase, whose coding sequences are downstream of the beta-galactosidase coding region, are inhibited to a lesser degree, 80% and 55%, respectively. The generation of anti-mRNA that can be targeted to inhibit a single species of mRNA molecule within cells provides a potent mechanism by which specific transcripts can be translationally inactivated. This can be used to determine the function of proteins as well as to select cloned genes in a single rapid and convenient step.
构建了一种质粒，用于在噬菌体λ PL启动子的控制下生成与β-半乳糖苷酶mRNA互补的RNA。当产生这种反义mRNA时，β-半乳糖苷酶的合成受到显著抑制（98%）。半乳糖苷通透酶和转乙酰酶的合成，其编码序列位于β-半乳糖苷酶编码区域的下游，分别受到80%和55%程度较轻的抑制。能够靶向抑制细胞内单一mRNA分子种类的反义mRNA的产生，提供了一种有效的机制，通过该机制特定转录本可以被翻译失活。这可用于确定蛋白质的功能，以及在一个快速便捷的步骤中筛选克隆基因。
