Identification,mapping and cloning of the thymidine kinase gene of fish lymphocystis disease virus |
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| Affiliation: | 1. Laboratory of Biotechnology and Aquatic Genomics, Interdisciplinary Center for Aquaculture Research (INCAR), University of Concepción, P.O. Box 160-C, Concepción, Chile;2. Aquainnovo, Casilla 30B, Puerto Montt 5503032, Chile;3. Facultad de Ciencias Veterinarias y Pecuarias, Universidad de Chile, Av Santa Rosa 11735, La Pintana, Santiago 8820808, Chile;4. Facultad de Ciencias Agronómicas, Universidad de Chile, Av Santa Rosa 11315, La Pintana, Santiago 8820808, Chile;1. Sichuan Entry–Exit Inspection and Quarantine Bureau, Chengdu, China;2. Sichuan Animal Science Academy, Chengdu 610066, China;3. College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China;4. Shenzhen Entry–Exit Inspection and Quarantine Bureau, Shenzhen, China;5. Hainan Entry–Exit Inspection and Quarantine Bureau, Haikou, China |
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| Abstract: | The thymidine kinase (TK) gene of fish lymphocystis disease virus (FLDV) was identified by biochemical transformation of 3T3 TK negative (TK−) to 3T3 TK positive (TK+) cells using specific viral DNA sequences. DNA fragments of the viral genome used in this study were obtained from a defined gene library of FLDV genome containing the complete viral DNA sequences. The selection of the converted cells was carried out under the condition of the HAT selection procedure. The results of these experiments revealed that the EcoRI FLDV DNA fragment C (11.2 kbp; 0.611 to 0.718 map units) is able to transform 3T3 TK− to 3T3 TK+ cells. Additional experiments using the subclones of EcoRI DNA fragment C revealed that DNA sequences of 4.1 kbp size between the coordinates 0.669 to 0.718 of the FLDV genome possessed the ability for biochemical transformation, indicating that the TK gene locus is located in this particular region. |
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