Structural gene identification and mapping by DNA-mRNA hybrid-arrested cell-free translation.

作者：Brett M. Paterson, Bryan E. Roberts, Edward L. Kuff · 发表于：Proceedings of the National Academy of Sciences · 年份：1977 · DOI：10.1073/pnas.74.10.4370 · 被引用次数：544 · 研究领域：Animal Genetics and Reproduction、RNA and protein synthesis mechanisms、Viral Infectious Diseases and Gene Expression in Insects

We present a simple method for directly correlating structural gene sequences in DNA with their corresponding mRNAs. This is based upon the fact that mRNA hybridized with its complementary DNA will not direct the cell-free synthesis of a complete polypeptide. Full translational activity of the mRNA is recovered upon the heat melting of the hybrid. Utilizing the rabbit beta globin clone PbetaG1, we demonstrate the application of hybrid-arrested translation for the identification of structural gene sequences within recombinant DNA molecules. In addition, the method is used to locate and order precisely several adenovirus 2 polypeptides within the viral genome.