title: Structural gene identification and mapping by DNA-mRNA hybrid-arrested cell-free translation.
abstract: 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.
mesh_terms: D000818:Animals; D001483:Base Sequence; D002474:Cell-Free System; D002874:Chromosome Mapping; D004247:DNA / Q000235:genetics*; D004274:DNA, Recombinant; D005796:Genes; D009693:Nucleic Acid Hybridization; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics*; D011817:Rabbits
chemical_list: D004274:DNA, Recombinant; D012333:RNA, Messenger; D004247:DNA
references.citation: Cell. 1977 Apr;10(4):571-85
references.pmid: 558827
references.citation: Biochim Biophys Acta. 1976 Dec 13;454(3):587-91
references.pmid: 999917
references.citation: Virology. 1977 May 15;78(2):522-36
references.pmid: 867816
references.citation: Cell. 1976 Jun;8(2):163-82
references.pmid: 61066
references.citation: Nucleic Acids Res. 1977;4(5):1539-52
references.pmid: 19730
references.citation: Biochem Biophys Res Commun. 1977 Apr 25;75(4):1127-32
references.pmid: 324472
references.citation: Proc Natl Acad Sci U S A. 1977 Aug;74(8):3171-5
references.pmid: 269380
references.citation: Cell. 1977 Feb;10(2):265-73
references.pmid: 837449
references.citation: Eur J Biochem. 1976 AUG 1;67(1):247-56
references.pmid: 823012
references.citation: Proc Natl Acad Sci U S A. 1976 Jul;73(7):2294-8
references.pmid: 781674
references.citation: Proc Natl Acad Sci U S A. 1975 Apr;72(4):1344-8
references.pmid: 1093178
references.citation: Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408-12
references.pmid: 4504350
references.citation: Proc Natl Acad Sci U S A. 1973 Aug;70(8):2330-4
references.pmid: 4525168
references.citation: Cell. 1976 Feb;7(2):279-88
references.pmid: 60178
references.citation: Methods Enzymol. 1974;30:709-23
references.pmid: 4369395
references.citation: J Mol Biol. 1975 May 5;94(1):71-86
references.pmid: 806696
references.citation: Proc Natl Acad Sci U S A. 1974 Jul;71(7):2756-60
references.pmid: 4527690
references.citation: J Mol Biol. 1975 Jan 5;91(1):121-3
references.pmid: 171410
references.citation: Eur J Biochem. 1974 Jul 1;46(1):83-8
references.pmid: 4850204
references.citation: Cold Spring Harb Symp Quant Biol. 1975;39 Pt 1:457-74
references.pmid: 1057474
references.citation: Biochemistry. 1975 Aug 26;14(17):3787-94
references.pmid: 1174504
references.citation: Nature. 1970 Aug 15;227(5259):680-5
references.pmid: 5432063
references.citation: J Biol Chem. 1969 Aug 25;244(16):4406-12
references.pmid: 5806584