Umene K
Department of Virology, Faculty of Medicine, Kyushu University 60, Fukuoka, Japan.
J Virol. 1994 Jul;68(7):4377-83. doi: 10.1128/JVI.68.7.4377-4383.1994.
DNA fragments corresponding to the unit-length a sequence of herpes simplex virus type 1 (HSV-1) were identified in HSV-1 DNA preparations extracted by the method of Hirt. The DNA fragments were molecularly cloned, and nucleotide sequences were determined. Most termini of the fragments were at sites on DR1 corresponding to the termini of linear HSV-1 DNA generated by the cleavage-packaging system. In one-step growth experiments, DNA fragments of the unit-length a sequence appeared simultaneously with the termini of linear HSV-1 DNAs produced by cleavage of circular and concatemeric DNAs. Therefore, excision of the unit-length a sequence appeared closely related to the cleavage-packaging system. Termini of the excised DNA fragments of the variant a sequence with two DR2 arrays varied on the L-component side, while termini on the S-component side were at the site on DR1 corresponding to the authentic cleavage site. It is thus assumed that the cleavage-packaging system functions adequately on the DR1 second distal from the S component, and cleavages of other DR1 are rare and less accurate. If this notion is tenable, then most termini on the S-component side of the excised DNA fragments are derived from the second DR1 properly cleaved and should be constant, while termini on the L-component side are from regions on and around the DR1 third distal from the S component and may be variable. Cleavage of DR1 is likely to be affected by the topological relationship with the S component.
在通过赫特方法提取的单纯疱疹病毒1型(HSV-1)DNA制剂中,鉴定出了与HSV-1单位长度a序列相对应的DNA片段。这些DNA片段进行了分子克隆,并测定了核苷酸序列。大多数片段的末端位于DR1上与切割包装系统产生的线性HSV-1 DNA末端相对应的位点。在一步生长实验中,单位长度a序列的DNA片段与通过环状和串联DNA切割产生的线性HSV-1 DNA末端同时出现。因此,单位长度a序列的切除似乎与切割包装系统密切相关。具有两个DR2阵列的变体a序列的切除DNA片段的末端在L组分一侧有所不同,而S组分一侧的末端位于DR1上与真实切割位点相对应的位点。因此可以推测,切割包装系统在距S组分第二远的DR1上能充分发挥作用,而其他DR1的切割很少且不太准确。如果这个观点成立,那么切除的DNA片段S组分一侧的大多数末端来自正确切割的第二个DR1,应该是恒定的,而L组分一侧的末端来自距S组分第三远的DR1及其周围区域,可能是可变的。DR1的切割可能会受到与S组分拓扑关系的影响。