Watanabe S, Temin H M
Proc Natl Acad Sci U S A. 1982 Oct;79(19):5986-90. doi: 10.1073/pnas.79.19.5986.
The minimal cis-acting sequences outside the long terminal repeat (LTR) required for formation of an infectious retrovirus cloning vector were determined with recombinants of spleen necrosis virus (SNV) DNA and herpes simplex virus type 1 thymidine kinase gene. The 3' end of SNV DNA was removed to within 40 base pairs (bp) from the 3' LTR with only a 2-fold effect on the recovery of infectious recombinant virus. However, when the 5' end of SNV DNA was removed to within 100 bp from the 5' LTR, infectious recombinant virus was not recovered. Deletion mutants constructed around this latter region showed that nucleotides between 100 and 285 bp from the 5' LTR are necessary for encapsidation of genomic viral RNA. We call this region required for encapsidation E.
利用脾坏死病毒(SNV)DNA与单纯疱疹病毒1型胸苷激酶基因的重组体,确定了形成感染性逆转录病毒克隆载体所需的长末端重复序列(LTR)以外的最小顺式作用序列。将SNV DNA的3'末端去除至距3' LTR 40个碱基对(bp)以内,对感染性重组病毒的回收仅产生2倍的影响。然而,当将SNV DNA的5'末端去除至距5' LTR 100 bp以内时,未回收感染性重组病毒。围绕后一区域构建的缺失突变体表明,距5' LTR 100至285 bp之间的核苷酸对于基因组病毒RNA的包装是必需的。我们将这个包装所需区域称为E区。