Wu M, Chari S, Yanchis T, Mergia A
Department of Pathobiology, College of Veterinary Medicine, University of Florida, Gainesville 32610, USA.
J Virol. 1998 Apr;72(4):3451-4. doi: 10.1128/JVI.72.4.3451-3454.1998.
We have constructed a series of vectors based on simian foamy virus type 1 (SFV-1) to define the minimum cis-acting elements required for gene transfer. To characterize these vectors, we inserted the coding sequence of the bacterial lacZ gene linked to the cytomegalovirus immediate-early gene promoter. Introduction of a deletion mutation in the leader region between the 5' long terminal repeat and the start of the gag gene at position 1659 to 1694 completely abrogated gene transfer by the SFV-1 vector. Deletion of 39 nucleotides from position 1692 to 1731 in the leader region resulted in a significant reduction in the transducing-particle titer. Furthermore, we have identified a second cis-acting element located at the 3' end of the pol gene between position 6486 and 6975 to be critical for SFV-1 vector transduction. These results identify the two important cis-acting elements required for SFV-1 vector construction, and the finding of a cis-acting element in the pol gene is unique among retroviruses.
我们构建了一系列基于1型猿猴泡沫病毒（SFV-1）的载体，以确定基因转移所需的最小顺式作用元件。为了表征这些载体，我们插入了与巨细胞病毒立即早期基因启动子相连的细菌lacZ基因的编码序列。在5'长末端重复序列与gag基因起始位点之间的前导区第1659至1694位引入缺失突变，完全消除了SFV-1载体的基因转移。在前导区第1692至1731位缺失39个核苷酸导致转导颗粒滴度显著降低。此外，我们确定了位于pol基因3'端第6486至6975位的第二个顺式作用元件对SFV-1载体转导至关重要。这些结果确定了构建SFV-1载体所需的两个重要顺式作用元件，并且在pol基因中发现顺式作用元件在逆转录病毒中是独特的。
