Cole M D, Ono M, Huang R C
J Virol. 1982 Apr;42(1):123-30. doi: 10.1128/JVI.42.1.123-130.1982.
Adjacent intracisternal A-particle (IAP) genes were identified in two different recombinant DNA clones, gamma 81 and gamma 19. In clone gamma 81, the most common form of IAP gene was separated by 5.3 kilobases from another IAP gene that had two apparent internal deletions. The two genes were in a head-to-tail configuration. In clone gamma 19, two different types of IAP genes were separated by less than 0.5 kilobase. Blot hybridization analysis of mouse DNA demonstrated that the DNA sequence found in clone gamma 81 is identical to the in vivo configuration. Using isolated DNA fragments from clone gamma 19, we mapped the boundaries of the IAP RNA by S1 digestion of RNA-DNA hybrids and by cDNA extension. With these techniques, both the 5' end and the 3' end of the IAP RNA in two different plasmacytomas (MOPC 315 and TEPC 15) were shown to fall within the long terminal direct repeat of the IAP gene. The fragment sizes generated by S1 digestion of IAP RNAs isolated from the two tumor lines were found to differ, indicating that different IAP genes may be transcribed in these two plasmacytomas.
在两个不同的重组DNA克隆γ81和γ19中鉴定出相邻的脑池内A颗粒(IAP)基因。在克隆γ81中,最常见形式的IAP基因与另一个有两个明显内部缺失的IAP基因相隔5.3千碱基。这两个基因呈头对头排列。在克隆γ19中,两种不同类型的IAP基因相隔不到0.5千碱基。对小鼠DNA的印迹杂交分析表明,克隆γ81中发现的DNA序列与体内结构相同。利用从克隆γ19中分离的DNA片段,我们通过RNA-DNA杂交体的S1消化和cDNA延伸来绘制IAP RNA的边界。通过这些技术,在两种不同的浆细胞瘤(MOPC 315和TEPC 15)中,IAP RNA的5'端和3'端均显示位于IAP基因的长末端直接重复序列内。发现从这两个肿瘤系中分离的IAP RNA经S1消化产生的片段大小不同,表明在这两种浆细胞瘤中可能转录不同的IAP基因。