Cloning and nucleotide sequence analysis of complementary deoxyribonucleic acid for bovine preproinsulin |
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Authors: | J D'Agostino M A Younes J W White P K Besch J B Field M L Frazier |
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Institution: | Department of Obstetrics and Gynecology, Baylor College of Medicine, Houston, Texas 77030. |
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Abstract: | A cDNA library was prepared from poly(A+) RNA isolated from fetal bovine pancreas. Bacterial colonies were screened for sequences homologous to a rat preproinsulin I cDNA probe. Ten positive clones were selected at random and further studied. Northern blot analyses revealed that seven of these clones hybridized to a single RNA species, of approximately 400 nucleotides. Sequence analysis of one of these clones (pbI2885) revealed the entire structural region of bovine preproinsulin mRNA including a 72 nucleotide region encoding a signal peptide enriched in hydrophobic residues. The overall nucleotide homology between bovine and human preproinsulin mRNA was 76% for the preregion, 89% for the A chain, 83% for the B chain, and 68% for the C peptide (including a 15 nucleotide deletion). |
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