期刊
JOURNAL OF VETERINARY MEDICAL SCIENCE
卷 63, 期 12, 页码 1297-1302出版社
JAPAN SOC VET SCI
DOI: 10.1292/jvms.63.1297
关键词
anti-microtubule drug; beta-tubulin; canine; isotype; ribonuclease protection assay
The structure of isotype-specific regions of classes I, II, III, IVa and lVb of canine beta -tubulin was characterized by 3'-RACE and the expression of these isotypes in canine tissues was examined by ribonuclease protection assay (RPA). Furthermore, a malignant mammary tumor-derived osteosarcoma-like cell line was established and the altered expression of beta -tubulin isotypes in taxol-resistant sublines was analyzed. The deduced amino acid sequences in isotype-specific regions corresponding to classes I, II and lVb were identical to those of humans and mice, but those in classes III and IVa showed slight differences among species. RPA revealed that classes I and IVb were widely distributed, but classes II, III and IVa were restricted to the brain. Because RPA could clearly distinguish the expression of class IVa from that of class IVb, it was thought to be more useful than northern blot for analysis of beta -tubulin isotype expression. In vitro, taxol-resistant sublines displayed a significant increase in class IVa as compared with taxol-sensitive cells, suggesting that altered expression of class IVa was associated with taxol resistance in these cell lines.
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