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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Mach, B. Rougeon, F. |
| Abstract | Two approaches have been explored for the synthesis of double-stranded DNA from single-stranded DNA template complementary to rabbit 9S globin mRNA (cDNA). (i) cDNA was elongated with dCMP or dTMP homopolymeric tracts using terminal deoxynucleotidyltransferase (EC 2.7.7.31; nucleosidetriphosphate:DNA deoxynucleotidylexotransferase). cDNA-dC, in the presence of an oligo(dG)10 primer, was an efficient template with either DNA polymerase of Escherichia coli (EC 2.7.7.7; deoxynucleosidetriphosphate:DNA deoxynucleotidyltransferase) or RNA-directed DNA polymerase of avian myeloblastosis virus. cDNA-dT [ with an oligo(dA)10 primer] functioned as template only with E. coli polymerase. (ii) cDNA, without homopolymeric tails, was also efficiently copied in the absence of oligonucleotide primer, by DNA polymerase of avian myeloblastosis virus or of E. coli. The product of the reaction consisted of long hairpin molecules which could be converted into DNA duplex (melting temperature, 93 degrees) by digestion with single-strand nuclease S1. The data indicate that a loop structure on the 3' end of cDNA allowed DNA synthesis to take place by a 'self-priming' mechanism. Some of the double-stranded DNA synthesized corresponded to the entire sequence of the 9S mRNA template. The synthesis of full-length double-stranded DNA from mouse globin mRNA and immunoglobulin light chain mRNA is also discussed. |
| ISSN | 00278424 |
| e-ISSN | 10916490 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Issue Number | 10 |
| Volume Number | 73 |
| Language | English |
| Publisher | National Academy of Sciences |
| Publisher Date | 1976-12-01 |
| Publisher Place | United States |
| Access Restriction | Open |
| Subject Keyword | DNA Metabolism Genes Globins RNA, Messenger Avian Myeloblastosis Virus Enzymology DNA Nucleotidyltransferases DNA, Single-Stranded Biosynthesis DNA-Directed DNA Polymerase Dactinomycin Pharmacology Escherichia Coli Immunoglobulin Light Chains Molecular Weight RNA-Directed DNA Polymerase Templates, Genetic Multidisciplinary |
| Content Type | Text |
| Resource Type | Article |
| Subject | Multidisciplinary |
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