DNA sequence analysis of spontaneoushprt mutations arising in vivo in cynomolgus monkey T-lymphocytes
| العنوان: | DNA sequence analysis of spontaneoushprt mutations arising in vivo in cynomolgus monkey T-lymphocytes |
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| المؤلفون: | P.R. Harbach, D.M. Zimmer, C.S. Aaron, S. S. Mattano, Y. Wang |
| المصدر: | Environmental and Molecular Mutagenesis. 26:218-225 |
| بيانات النشر: | Wiley, 1995. |
| سنة النشر: | 1995 |
| مصطلحات موضوعية: | Male, Hypoxanthine Phosphoribosyltransferase, Epidemiology, Sequence analysis, T-Lymphocytes, Health, Toxicology and Mutagenesis, Molecular Sequence Data, Mutant, Animals, Wild, Biology, Polymerase Chain Reaction, Exon, Complementary DNA, Animals, Humans, Coding region, RNA, Messenger, Gene, Genetics (clinical), DNA Primers, Sequence Deletion, Genetics, Base Composition, Base Sequence, Nucleic acid sequence, Intron, DNA, Exons, Molecular biology, Introns, Clone Cells, Macaca fascicularis, DNA Transposable Elements, Female, Artifacts |
| الوصف: | To study the mechanisms of mutagenesis in vivo, we analyzed mutations at the hypoxanthine phosphoribosyl transferase (hprt) locus using cDNA from cynomolgus monkey T-lymphocytes. In the present study, the spectrum of spontaneous hprt mutations arising in vivo in wild-caught cynomolgus monkey peripheral T-lymphocytes is described. Cells were isolated from peripheral blood, and mutant clones were selected in 6-thioguanine, propagated, and stored frozen. cDNA was copied from hprt mRNA from a lysate of 7,000 to 20,000 cells. A 780-base-pairs (bp) region including the coding region was amplified by polymerase chain reaction and directly sequenced. We sequenced 40 spontaneous mutants from 11 monkeys. Of these 40 clones, 23 (57%) had base-pair substitutions, 11 (28%) had small (20 bp) deletions and/or insertions, and 6 (15%) had large (20 bp) deletions and/or insertions. Of the 23 base substitutions, 13 were transitions (11 G:C--A:T, 1 A:T--G:C, and 1 tandem TT--CC) and 10 were transversions (3 G:C--T:A, 3 G:C--C:G, 2 A:T--T:A, 2 A:T--C:G). Bases 209 and 617 were apparent substitution hotspots, which have also been observed as hotspots in human hprt. In 2 clones with large insertions, the inserted bases were of intronic origin. One of these lost 272 bp from exons 2-3 and contained a 93-bp insertion from the middle of intron 3. Two clones with small deletions and 5 clones with large deletions or insertions (7/40 or 17.5%) could be splice mutants. |
| تدمد: | 1098-2280 0893-6692 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::97e26d0f983f386bbc8109e6a8c23c0cTest https://doi.org/10.1002/em.2850260306Test |
| حقوق: | CLOSED |
| رقم الانضمام: | edsair.doi.dedup.....97e26d0f983f386bbc8109e6a8c23c0c |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 10982280 08936692 |
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