Tiollais P, Perricaudet M, Pettersson U, Philipson L
Gene. 1976;1(1):49-63. doi: 10.1016/0378-1119(76)90006-8.
Hybrid genomes of bacteriophage lambda with integrated fragments of adenovirus type 2 (Ad2) DNA have been constructed in vitro and propagated in E. coli. DNA from a derivative of bacteriophage lambdaplac5 (Rambach and Tiollais, 1974) was used as a vector. The two fragments of the vector DNA contain all the essential genes for the replication of the lambda DNA but are too short to be encapsidated. Insertion of DNA is therefore essential for plaque formation which constitutes a selection method for phages containing hybrid genomes. Fragments EcoRI-B and EcoRI-F of Ad2 DNA were purified, and separately ligated with the vector fragments. Clones of hybrid phage could readily be isolated. Two clones of hybrid phage containing fragment EcoRI-B inserted in opposite directions were used to study the transcription of adenovirus-specific sequences. Hybridization experiments showed that transcripts from both strands of fragment Ad2-Eco RI-B could be detected and that transcription probably was controlled by the "early" leftward and the "late" rightward promoters on the lambda genome. No polypeptides specified by the adenovirus fragment have so far been identified.
已经在体外构建了带有2型腺病毒(Ad2)DNA整合片段的噬菌体λ杂交基因组,并在大肠杆菌中进行增殖。来自噬菌体λplac5衍生物(Rambach和Tiollais,1974)的DNA用作载体。载体DNA的两个片段包含λDNA复制所需的所有必需基因,但太短而无法被包装。因此,DNA的插入对于噬菌斑形成至关重要,这构成了一种针对含有杂交基因组的噬菌体的选择方法。纯化了Ad2 DNA的EcoRI - B和EcoRI - F片段,并分别与载体片段连接。杂交噬菌体的克隆很容易分离出来。使用两个含有以相反方向插入的EcoRI - B片段的杂交噬菌体克隆来研究腺病毒特异性序列的转录。杂交实验表明,可以检测到片段Ad2 - Eco RI - B两条链的转录本,并且转录可能受λ基因组上的“早期”向左和“晚期”向右启动子控制。到目前为止,尚未鉴定出腺病毒片段所指定的多肽。