Cloning and nucleotide sequence of a mouse erythrocyte beta-spectrin cDNA
| Autor: | Steven Goodman, Keith Krebs, Pekka Laurila, Livia Cioè, Pacifico Meo, Peter J. Curtis |
|---|---|
| Rok vydání: | 1987 |
| Předmět: |
Male
Erythrocytes Molecular Sequence Data Immunology Biology Biochemistry Mice Nucleic acid thermodynamics Complementary DNA Animals RNA Messenger Cloning Molecular Peptide sequence Southern blot chemistry.chemical_classification Base Sequence Nucleic acid sequence Spectrin RNA DNA Cell Biology Hematology Molecular biology Amino acid genomic DNA chemistry Genetic Code Immunologic Techniques Rabbits |
| Zdroj: | Blood. 70:915-920 |
| ISSN: | 1528-0020 0006-4971 |
| DOI: | 10.1182/blood.v70.4.915.915 |
| Popis: | A rabbit monospecific antibody for mouse beta-spectrin was used to screen a mouse anemic spleen cDNA expression library. A mouse beta- spectrin cDNA clone was isolated and identified by its ability to make mouse beta-spectrin-like antigens in Escherichia coli. This clone was used to probe total RNA from various mouse tissues. Anemic spleen RNA showed two strongly hybridizing RNA species of approximately 6 and 8 kb. Two very faintly hybridizing bands of about 6 kb and 10 kb could also be seen in total mouse brain RNA. All of these bands could be detected after hybridization under both stringent and nonstringent conditions. This suggests that erythroid beta-spectrin may also be expressed in the brain. No bands could be detected in kidney, liver, or spleen RNA. Southern blot analysis of mouse genomic DNA showed a single hybridizing band after digestion with several restriction endonucleases even under nonstringent conditions. Nucleotide sequencing of the cDNA insert revealed almost complete identity between the N-terminus of the deduced amino acid sequence of the cDNA clone and the C-terminal 15 amino acids of a peptide derived from the beta-8 repeat unit of human erythrocyte beta-spectrin. The deduced amino acid sequence contained most of the conserved amino acids characteristic of the 106 amino acid repeat unit first found in human alpha-spectrin and thus provides the first evidence for a complete 106 amino acid repeat unit structure in beta-spectrin. |
| Databáze: | OpenAIRE |
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