The synthesis of high yields of full-length reverse transcripts of globin mRNA
- 1 January 1977
- journal article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 4 (10) , 3455-3471
- https://doi.org/10.1093/nar/4.10.3455
Abstract
Conditions have been determined under which reverse transcriptase catalyzes the synthesis of the high yields of full length complementary deoxyribonucleic acid (cDNA). These conditions depend not only on the cencentration of deoxynucleoside triphosphates (1) but also on the concentration of reverse transcriptase. An analysis of the kinetics of cDNA synthesis and the size of cDNA synthesized as a function of time under different conditions indicates that the mechanism of action of reverse transcriptase is partially distributive. This accounts for the necessity of a high enzyme concentration to obtain high yields of full length cDNA. Additional experiments indicate that the yield of cDNA is limited by the fact that the template mRNA is rapidly inactivated. This is most likely due to the fact that the product cDNA is hydrogen bonded to the template mRNA during synthesis.Keywords
This publication has 23 references indexed in Scilit:
- Number and distribution of polyadenylated RNA sequences in yeastCell, 1977
- Chain length determination of small double- and single-stranded DNA molecules by polyacrylamide gel electrophoresisBiochemistry, 1975
- The steady state kinetic parameters and non-processivity of Escherichia coli deoxyribonucleic acid polymerase IJournal of Biological Chemistry, 1975
- Sequence at the 3′ end of globin mRNA shows homology with immunoglobulin light chain mRNANature, 1974
- Nucleotide Sequence Analysis of RNA Synthesized from Rabbit Globin Complementary DNAProceedings of the National Academy of Sciences, 1974
- Nucleotide Sequences of Human Globin Messenger RNAProceedings of the National Academy of Sciences, 1974
- Kinetic studies of gene frequency: I. Use of a DNA copy of reticulocyte 9 S RNA to estimate globin gene dosage in mouse tissuesJournal of Molecular Biology, 1974
- Demonstration of Globin Messenger Sequences in Giant Nuclear Precursors of Messenger RNA of Avian ErythroblastsProceedings of the National Academy of Sciences, 1973
- Isolation of poly(adenylic acid)-rich ribonucleic acid from mouse myeloma and synthesis of complementary deoxyribonucleic acidBiochemistry, 1973
- Primer Requirement and Template Specificity of the DNA Polymerase of RNA Tumor VirusesProceedings of the National Academy of Sciences, 1971