Miyazawa T, Tomonaga K, Kawaguchi Y, Kohmoto M, Inoshima Y, Maeda K, Mikami T, Maedadel K [corrected to Maeda K ]
Department of Veterinary Microbiology, Faculty of Agriculture, University of Tokyo, Japan.
Arch Virol. 1994;139(1-2):37-48. doi: 10.1007/BF01309453.
An oligonucleotide containing multiple AP-1 binding sites was introduced into the regulatory sequence in the long terminal repeat (LTR) of feline immunodeficiency virus (FIV). Chloramphenicol acetyltransferase assay revealed that basal promoter activity of the mutated LTR was higher than that of the wild-type LTR in Crandell feline kidney (CRFK) cells. The mutated LTR was introduced into an infectious molecular clone of FIV and the clone was transfected into CRFK cells. The virus production of the mutant in the cells was as high as that of the wild-type when determined by the reverse transcriptase activity assay. The growth of the mutant virus obtained from the transfected CRFK cells was examined in feline T lymphoblastoid cell lines (MYA-1 and FeL-039 cells) and primary feline peripheral blood mononuclear cells (fPBMCs). The growth was delayed when compared with that of the wild-type virus in all the cells used. Upon examination by polymerase chain reaction, the length of the LTR of the mutant virus was shortened in both MYA-1 cells and fPBMCs. Sequence analysis revealed that the insertion was completely deleted 39 days after infection in the MYA-1 cells.
将一个含有多个AP-1结合位点的寡核苷酸导入猫免疫缺陷病毒（FIV）长末端重复序列（LTR）的调控序列中。氯霉素乙酰转移酶分析表明，在克兰德尔猫肾（CRFK）细胞中，突变LTR的基础启动子活性高于野生型LTR。将突变的LTR导入FIV的感染性分子克隆中，并将该克隆转染到CRFK细胞中。通过逆转录酶活性测定法测定，细胞中突变体的病毒产生量与野生型一样高。在猫T淋巴母细胞系（MYA-1和FeL-039细胞）和原代猫外周血单个核细胞（fPBMC）中检测了从转染的CRFK细胞获得的突变病毒的生长情况。与野生型病毒相比，在所有使用的细胞中，突变病毒的生长均延迟。通过聚合酶链反应检测，在MYA-1细胞和fPBMC中，突变病毒的LTR长度均缩短。序列分析显示，在MYA-1细胞感染39天后，插入片段完全缺失。
