Identification, mapping and cloning of the thymidine kinase gene of fish lymphocystis disease virus.
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.
Scholz
J
J
Institut für Medizinische Virologie der Universität Heidelberg, F.R.G.
Rösen-Wolff
A
A
Touray
M
M
Schnitzler
P
P
Darai
G
G
D000818:Animals; D004279:DNA, Viral / Q000235:genetics; D005347:Fibroblasts; D007497:Iridoviridae / Q000201:enzymology / Q000235:genetics*; D051379:Mice; D011994:Recombinant Proteins; D013937:Thymidine Kinase / Q000235:genetics*; D014162:Transfection; D014764:Viral Proteins / Q000235:genetics*
D004279:DNA, Viral; D011994:Recombinant Proteins; D014764:Viral Proteins; D013937:Thymidine Kinase