Cloning of bacteriophage fd gene 2 and construction of a plasmid dependent on fd gene 2 protein.

Meyer, Thomas F.; Geider, Klaus

doi:10.1073/pnas.78.9.5416

Cited by 18 publications

(7 citation statements)

References 25 publications

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“…Recently, gene 2 has been cloned into expression plasmids (141,206). These systems enable a large-scale production and isolation of biologically active g2p.…”

Section: Replication Proteinsmentioning

confidence: 99%

Ff coliphages: structural and functional relationships.

Rasched¹,

Oberer²

1986

Microbiological Reviews

105

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“…Recently, gene 2 has been cloned into expression plasmids (141,206). These systems enable a large-scale production and isolation of biologically active g2p.…”

Section: Replication Proteinsmentioning

confidence: 99%

Ff coliphages: structural and functional relationships.

Rasched¹,

Oberer²

1986

Microbiological Reviews

105

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“…It was reported previously that gene II of phage fd is sufficient for replication of plasmids carrying the intergenic region of phage fd (20). Derivatives of phasyl which were defective in the arp gene could replicate in a strain carrying the gene II product, but the arp protein did not complement gene II mutants of phage fd.…”

mentioning

confidence: 98%

Characterization of a phage-plasmid hybrid (phasyl) with two independent origins of replication isolated from Escherichia coli

1991

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The phage-plasmid hybrid phasyl can replicate as a phage in the presence of a filamentous phage of Escherichia coli (M13, fl, fd). The extragenic region of phasyl shows homology with the plus and the minus origins of filamentous phages. Insertion of a Cmr fragment into the plus origin or of a Kmr fragment into the minus origin resulted in a reduced transduction frequency, while insertion into other parts of the extragenic region did not. This suggests that phagelike replication of phasyl is mediated by an origin that coincides with the two homologous elements in the extragenic region. Autonomous replication of phasyl occurs from a second origin (oriA) that is located between positions 297 and 636. This fragment mediates replication if the Arp protein is supplied in trans. Arp is the only phage-encoded protein and is essential for plasmidlike replication. No sequence homology to other known origins was found. Phasyl derivatives with either one of the two origins inactivated can be rescued via the alternative replication mode, suggesting that the two replication pathways are independent.Phasyl is the smallest known naturally occurring replicon in Escherichia coli (1,282 bp) ( Fig. 1) (25). It is a phageplasmid hybrid (phasmid). For autonomous replication, the Arp protein (autonomous replication of phasyl) is essential. The Arp protein is the only gene product expressed by phasyl known so far. Any manipulation in this reading frame leads to the loss of autonomous replication capability (25). In the presence of a filamentous phage (M13, fd, fl), phasyl can replicate via the phage mode. Single-stranded DNA (ss-DNA) is packaged, and phage particles are secreted.The 424-bp extragenic region of phasyl shows two elements which are homologous by sequence and potential secondary structure with the origins for viral (plus)-and complementary (minus)-strand DNA synthesis of filamentous E. coli phages (Fig. 2). However, the order of plus-and minus-strand origin is reversed in phasyl when compared with the filamentous phages, and the degree of homology is lower. Also, the morphogenetic signal (domain A) and the A+T-rich enhancer region (domain F), which supposedly are essential features in the intergenic region of filamentous phages, are missing (4,5,10,16,25,29).It was reported previously that gene II of phage fd is sufficient for replication of plasmids carrying the intergenic region of phage fd (20). Derivatives of phasyl which were defective in the arp gene could replicate in a strain carrying the gene II product, but the arp protein did not complement gene II mutants of phage fd. Also, the rep helicase, which is required for the rolling circle replication of single-stranded phages (6), is not essential for autonomous replication of phasyl (25).From all these arguments, it appeared that two distinct mechanisms of replication are involved during the growth of phasyl as a plasmid or as defective phage. However, it was not known whether the identified origin in the extragenic region, which is similar to that of filamentous pha...

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“…A DNA fragment of 260 nucleotides was dissected from the fd genome and linked to a gene for resistance to kanamycin (NPT I) (Meyer & Geider, 1981). Because propagation of this artificial plasmid (pfd) depends on the presence of gene 2 protein in the same cell, the gene coding for this protein was inserted into a pBR plasmid and provided a sufficient supply of the viral replication protein.…”

Section: Plasmids Using Fd Replication Functionsmentioning

confidence: 99%

“…Viral strands are replicated in the rolling circle mode and circularized via cleavage of the single-stranded tails and strand sealing by gene 2 protein (Meyer & Geider, 1982). Gene 2 of filamentous bacteriophage has been cloned into pBR plasmids (Meyer & Geider, 1981), thus increasing the (Johnston et al, 1985). Plasmids having the replication origin of phage fd seem to stimulate the expression of cloned gene 2 (Meyer & Geider, 1981).…”

Section: Introductionmentioning

confidence: 99%

“…Gene 2 of filamentous bacteriophage has been cloned into pBR plasmids (Meyer & Geider, 1981), thus increasing the (Johnston et al, 1985). Plasmids having the replication origin of phage fd seem to stimulate the expression of cloned gene 2 (Meyer & Geider, 1981). Gene 5 protein prevents initiation of complementary strand synthesis by binding to the single-stranded tails of the rolling circles.…”

Section: Introductionmentioning

confidence: 99%

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