Denman R B
New York State Institute for Basic Research in Developmental Disabilities, Staten Island, New York 10314, USA.
Biochem Biophys Res Commun. 1997 Jan 3;230(1):226-31. doi: 10.1006/bbrc.1996.5926.
The behavior of ribozyme and antisense RNAs was analyzed in a coupled rabbit reticulocyte transcription translation system. Both ribozyme and antisense RNAs were efficiently produced and bound tightly to polyribosomes at 30 degrees C, but did not produce a protein product. Antisense and ribozyme RNA binding depended upon the presence of intact ribosomes, was specific since, plasmid DNA did not associate with either ribosomes or polyribosomes, and was temperature dependent. Ribozyme-specific mRNA cleavage in the coupled system was inferred from translation inhibition studies and was confirmed by primer extension analysis. Thus, ribozyme RNA can inhibit target protein production in the coupled transcription translation system by competing out cellular mRNAs and via targeted message degradation.
在兔网织红细胞转录翻译偶联系统中分析了核酶和反义RNA的行为。核酶和反义RNA均能高效产生，并在30℃时紧密结合到多核糖体上，但未产生蛋白质产物。反义RNA和核酶RNA的结合依赖于完整核糖体的存在，具有特异性，因为质粒DNA不与核糖体或多核糖体结合，且具有温度依赖性。从翻译抑制研究中推断出偶联系统中核酶特异性的mRNA切割，并通过引物延伸分析得到证实。因此，核酶RNA可通过竞争细胞内mRNA并通过靶向信息降解来抑制偶联转录翻译系统中靶蛋白的产生。
