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A region of the filamentous phage phi Lf genome that can support autonomous replication and miniphage production

N T Lin1, F S Wen, Y H Tseng

  • 1Department of Botany, National Chung Hsing University, Taichung, Taiwan, Republic of China.

Biochemical and Biophysical Research Communications
|January 5, 1996
PubMed

Insights

Researchers identified a key replication gene (gene II) in the Xanthomonas phage phi Lf. This discovery advances understanding of filamentous phage replication and DNA integration mechanisms.

Area of Science:

  • Molecular Biology
  • Virology
  • Bacteriology

Background:

  • Filamentous phages like phi Lf are important tools in molecular biology.
  • Understanding phage replication mechanisms is crucial for genetic engineering and phage therapy.

Purpose of the Study:

  • To characterize the replication initiation protein gene (gene II) of the Xanthomonas campestris pv. campestris filamentous phage phi Lf.
  • To investigate the regulatory elements involved in phi Lf replication and integration.

Main Methods:

  • Construction and maintenance of a phi Lf minireplicon in Xanthomonas cells.
  • Analysis of the open reading frame (ORF346) within the minireplicon.
  • Bioinformatic analysis of conserved protein motifs and DNA sequences.

Main Results:

  • A 2028-bp fragment of phi Lf DNA functioned as an autonomous minireplicon.
  • ORF346 encodes a 39,144 MW polypeptide with Rep protein consensus motifs, identified as phi Lf gene II.
  • Sequences resembling Ff phage intergenic regions, including IHF binding sites and an AT-rich integration core, were identified upstream and downstream of ORF346.

Conclusions:

  • ORF346 is the gene encoding the replication initiation protein (gene II) of phi Lf.
  • The identified upstream and downstream sequences suggest conserved mechanisms for replication and integration in filamentous phages.
  • The phi Lf minireplicon system provides a platform for studying phage replication and gene expression.

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