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

Abstract
A simple method is presented for directly correlating structural gene sequences in DNA with their corresponding mRNA. 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 P.beta.G1, the application of hybrid-arrested translation for the identification of structural gene sequences within recombinant DNA molecules is demonstrated. The method is also used to locate and order precisely several adenovirus 2 polypeptides within the viral genome.