Krause H, Wunderlich V, Uckert W
Department of Virology, Academy of Sciences of the German Democratic Republic, Berlin.
Virology. 1989 Nov;173(1):214-22. doi: 10.1016/0042-6822(89)90237-7.
Unintegrated circular proviral DNA of a type D retrovirus (PMFV) isolated from a permanent human cell line was molecularly cloned in the bacteriophage vector L47.1 and subcloned in the plasmid vector pGEM-2. A restriction endonuclease map of PMFV DNA was established using 10 different enzymes for single and multiple digestions of closed circular and cloned DNA molecules. By restriction endonuclease analysis cloned PMFV DNA represented full-length viral DNA with one long terminal repeat (LTR). The comparison of the physical map of cloned PMFV to those of other cloned type D retroviruses revealed closest homology to the map of retrovirus D/New England (pD398) and SAIDS retrovirus type 1 (SRV-1). The relatedness of PMFV to further type D retroviruses (Mason-Pfizer monkey virus, MPMV; SAIDS retrovirus type 2, SRV-2) was also demonstrated by cross-hybridization of cloned DNAs under different stringencies (i) using full-length genomic probes of PMFV, MPMV, and SRV-2 and (ii) by DNA sequence analysis of regions of the group specific antigen (gag) protease (prt), polymerase (pol), and envelope (env) genes.
从一个永久性人类细胞系中分离出的D型逆转录病毒（PMFV）的未整合环状前病毒DNA在噬菌体载体L47.1中进行分子克隆，并在质粒载体pGEM-2中进行亚克隆。使用10种不同的酶对封闭环状和克隆的DNA分子进行单酶切和多酶切，建立了PMFV DNA的限制性内切酶图谱。通过限制性内切酶分析，克隆的PMFV DNA代表具有一个长末端重复序列（LTR）的全长病毒DNA。将克隆的PMFV物理图谱与其他克隆的D型逆转录病毒的图谱进行比较，发现与逆转录病毒D/新英格兰（pD398）和1型艾滋病相关逆转录病毒（SRV-1）的图谱具有最密切的同源性。通过在不同严格条件下（i）使用PMFV、MPMV和SRV-2的全长基因组探针以及（ii）对群特异性抗原（gag）蛋白酶（prt）、聚合酶（pol）和包膜（env）基因区域进行DNA序列分析，克隆DNA的交叉杂交也证明了PMFV与其他D型逆转录病毒（梅森-辉瑞猴病毒，MPMV；2型艾滋病相关逆转录病毒，SRV-2）的相关性。
