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丝状噬菌体IKe和Ff(M13、fd和f1)复制与包装相关DNA序列的比较

Comparison of the DNA sequences involved in replication and packaging of the filamentous phages IKe and Ff (M13, fd, and f1).

作者信息

Peeters B P, Schoenmakers J G, Konings R N

出版信息

DNA. 1987 Apr;6(2):139-47. doi: 10.1089/dna.1987.6.139.

Abstract

The product of gene II of the distantly related, filamentous, single-stranded DNA phages IKe and Ff (M13, fd, and f1) is the only phage-encoded protein that is required for the replication of their double-stranded replicative form DNA. With the aid of recombinant plasmids containing the origins of viral strand replication [(+)-origins] of both IKe and Ff, we demonstrated that initiation but not termination of viral strand replication by gene II protein is restricted to its cognate (+)-origin. If the (+)-origins of IKe and Ff are present in the same orientation, fusion origins are generated upon gene II protein-instructed replication as a result of initiation at one origin and termination at the other. These fusion origins are only functional in the presence of the gene II protein encoded by the phage from which the sequence lying at the 3' side of the gene II protein cleavage site is derived. The nucleotides that determine the specificity of the replication initiation process are located between positions +17 and +49, or +17 and +40, with respect to the gene II protein cleavage site of IKe and Ff, respectively. The DNA sequence that forms the recognition signal for cleavage by gene II protein is probably located within the sequence that starts 3 nucleotides before and terminates 17 nucleotides after the cleavage site. Efficient packaging by phage IKe of plasmid DNA strands that contain the morphogenetic signal of Ff, or vice versa, indicates that, despite their only partial homology, the morphogenetic signals of IKe and Ff are interchangeable.

摘要

远缘丝状单链DNA噬菌体IKe和Ff(M13、fd和f1)的基因II产物是其双链复制型DNA复制所必需的唯一噬菌体编码蛋白。借助含有IKe和Ff病毒链复制起点[(+)-起点]的重组质粒,我们证明基因II蛋白引发病毒链复制而非终止复制仅限于其同源(+)-起点。如果IKe和Ff的(+)-起点以相同方向存在,那么在基因II蛋白指导的复制过程中,由于在一个起点起始复制并在另一个起点终止复制,会产生融合起点。这些融合起点仅在存在由噬菌体编码的基因II蛋白时才起作用,该噬菌体的基因II蛋白切割位点3'侧的序列即来源于此。相对于IKe和Ff的基因II蛋白切割位点,决定复制起始过程特异性的核苷酸分别位于+17至+49位或+17至+40位之间。形成基因II蛋白切割识别信号的DNA序列可能位于切割位点前3个核苷酸起始、切割位点后17个核苷酸终止的序列内。噬菌体IKe能有效包装含有Ff形态发生信号的质粒DNA链,反之亦然,这表明尽管IKe和Ff的形态发生信号仅有部分同源性,但它们是可互换的。

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