Wu Y, Zhang D Y, Kramer F R
Department of Molecular Genetics, Public Health Research Institute, New York, NY 10016.
Proc Natl Acad Sci U S A. 1992 Dec 15;89(24):11769-73. doi: 10.1073/pnas.89.24.11769.
RNA molecules were prepared that consisted of an mRNA encoding chloramphenicol acetyltransferase embedded within the sequence of midivariant RNA, which is a template for the RNA-directed RNA polymerase Q beta replicase. These recombinant RNAs were shown to be bifunctional: they are amplified exponentially by incubation with Q beta replicase, and the replicated RNA serves as template for the cell-free synthesis of enzymatically active chloramphenicol acetyltransferase. The availability of amplifiable mRNAs will enable relatively large amounts of protein to be synthesized in vitro.
制备了RNA分子，其由编码氯霉素乙酰转移酶的mRNA组成，该mRNA嵌入中变体RNA序列中，中变体RNA是RNA指导的RNA聚合酶Qβ复制酶的模板。这些重组RNA被证明具有双功能：通过与Qβ复制酶孵育可进行指数扩增，并且复制的RNA可作为无细胞合成具有酶活性的氯霉素乙酰转移酶的模板。可扩增mRNA的可用性将使体外合成相对大量的蛋白质成为可能。
