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源自日本鹌鹑细胞系QT35的胸苷激酶缺陷型禽成纤维细胞系的建立与鉴定

Establishment and characterization of a thymidine kinase deficient avian fibroblast cell line derived from a Japanese quail cell line, QT35.

作者信息

Niikura M, Narita T, Mikami T

机构信息

Department of Epizootiology, Faculty of Veterinary Medicine, Hokkaido University, Sapporo, Japan.

出版信息

J Vet Med Sci. 1991 Jun;53(3):439-46. doi: 10.1292/jvms.53.439.

Abstract

An avian thymidine kinase deficient (TK-) fibroblast cell line (QTTK-) was established from a Japanese quail cell line, QT35, and characterized the biological properties. QTTK- could grow in the presence of 5-bromo-2'-deoxyuridine (BUdR, 100 micrograms/ml) and not in the growth medium with hypoxanthine-aminopterin-thymidine. Compared to QT35 cells, the 3H-thymidine incorporation of the QTTK- cells and the TK activity of the cell extract significantly decreased to 0.3% and 0.5%, respectively. In the thymidylate synthetase activity, the karyotype, and the sensitivities to either fowlpox virus (FPV) or herpesvirus of turkeys (HVT), OTTK- cells were similar to the parental QT35 cells. Since QTTK- cells were permissive to FPV and HVT infections and these viruses could not grow in the presence of 50 to 75 micrograms/ml of BUdR, QTTK- cells may be useful for the construction of recombinant FPV and HVT that have foreign genes within the TK gene of the virus genome.

摘要

从日本鹌鹑细胞系QT35建立了一种禽胸苷激酶缺陷(TK-)成纤维细胞系(QTTK-),并对其生物学特性进行了表征。QTTK-可以在5-溴-2'-脱氧尿苷(BUdR,100微克/毫升)存在的情况下生长,而不能在含有次黄嘌呤-氨基蝶呤-胸腺嘧啶核苷的生长培养基中生长。与QT35细胞相比,QTTK-细胞的3H-胸腺嘧啶核苷掺入量和细胞提取物的TK活性分别显著降低至0.3%和0.5%。在胸苷酸合成酶活性、核型以及对禽痘病毒(FPV)或火鸡疱疹病毒(HVT)的敏感性方面,QTTK-细胞与亲代QT35细胞相似。由于QTTK-细胞允许FPV和HVT感染,并且这些病毒在50至75微克/毫升的BUdR存在下不能生长,QTTK-细胞可能有助于构建在病毒基因组的TK基因内含有外源基因的重组FPV和HVT。

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