Abstract
Sequences of human beta-globin mRNA were determined by analysis of complementary DNA. beta-mRNA was transcribed into double-stranded cDNA by RNA-dependent DNA polymerase. cDNA was cut by restriction endonucleases and the fragments were terminally labeled by means of polynucleotide kinase and [gamma-32P]ATP. After purification, fragments were degraded by snake venom phosphodiesterase. Alternatively single-stranded [32P]cDNA was prepared by transcription in the presence of [alpha-32P]dCTP and actinomycin D; the product was digested by endonuclease IV and degraded by snake venom phosphodiesterase. cDNA tracts obtained by both labeling methods enabled us to construct a sequence for the translated and 3'-terminal untranslated regions of human beta-mRNA.
Publication types
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Research Support, U.S. Gov't, Non-P.H.S.
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Anemia, Sickle Cell
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Base Sequence
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DNA Restriction Enzymes
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DNA*
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Dactinomycin
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Deoxyribonucleases
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Endonucleases
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Globins / biosynthesis*
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Humans
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Oligodeoxyribonucleotides
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Phosphoric Diester Hydrolases
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Polynucleotide 5'-Hydroxyl-Kinase
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Protein Biosynthesis
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RNA, Messenger* / metabolism
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RNA-Directed DNA Polymerase
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Thalassemia
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Transcription, Genetic
Substances
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Oligodeoxyribonucleotides
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RNA, Messenger
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Dactinomycin
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Globins
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DNA
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Polynucleotide 5'-Hydroxyl-Kinase
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RNA-Directed DNA Polymerase
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Deoxyribonucleases
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Endonucleases
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DNA Restriction Enzymes
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Phosphoric Diester Hydrolases
Associated data
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GENBANK/J00093
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GENBANK/J00094
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GENBANK/J00096
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GENBANK/J00158
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GENBANK/J00159
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GENBANK/J00160
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GENBANK/J00161
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GENBANK/J00162
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GENBANK/J00163
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GENBANK/J00164
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GENBANK/J00165
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GENBANK/J00166
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GENBANK/J00167
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GENBANK/J00168
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GENBANK/J00169
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GENBANK/J00170
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GENBANK/J00171
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GENBANK/J00172
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GENBANK/J00173
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GENBANK/J00174
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GENBANK/J00175
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GENBANK/J00177
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GENBANK/J00178
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GENBANK/J00179
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GENBANK/K01239
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GENBANK/K01890
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GENBANK/K02544
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GENBANK/M18047
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GENBANK/M19067
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GENBANK/X00423