Scholz J, Rösen-Wolff A, Touray M, Schnitzler P, Darai G
Institut für Medizinische Virologie der Universität Heidelberg, F.R.G.
Virus Res. 1988 Jan;9(1):63-72. doi: 10.1016/0168-1702(88)90050-0.
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.
利用鱼类淋巴囊肿病病毒（FLDV）的特定病毒DNA序列，通过将3T3胸苷激酶阴性（TK-）细胞生化转化为3T3胸苷激酶阳性（TK+）细胞，鉴定出了该病毒的胸苷激酶（TK）基因。本研究中使用的病毒基因组DNA片段，是从包含完整病毒DNA序列的FLDV基因组特定基因文库中获得的。转化细胞的筛选是在次黄嘌呤-氨基蝶呤-胸腺嘧啶核苷（HAT）选择程序的条件下进行的。这些实验结果表明，EcoRI FLDV DNA片段C（11.2千碱基对；0.611至0.718图谱单位）能够将3T3 TK-细胞转化为3T3 TK+细胞。使用EcoRI DNA片段C亚克隆的进一步实验表明，FLDV基因组坐标0.669至0.718之间大小为4.1千碱基对的DNA序列具有生化转化能力，这表明TK基因位点位于该特定区域。
