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Titolo:
Plasmids carrying cloned fragments of RF DNA from the filamentous phage phi Lf can be integrated into the host chromosome via site-specific integration and homologous recombination
Autore:
Lin, NT; Chang, RY; Lee, SJ; Tseng, YH;
Indirizzi:
Natl Chunghsing Univ, Inst Mol Biol, Taichung 402, Taiwan Natl Chunghsing Univ Taichung Taiwan 402 Mol Biol, Taichung 402, Taiwan Tzu Chi Univ, Dept Microbiol, Hualien 970, Taiwan Tzu Chi Univ Hualien Taiwan 970 niv, Dept Microbiol, Hualien 970, Taiwan Natl Dong Hwa Univ, Dept Life Sci, Hualien 974, Taiwan Natl Dong Hwa UnivHualien Taiwan 974 Dept Life Sci, Hualien 974, Taiwan
Titolo Testata:
MOLECULAR GENETICS AND GENOMICS
fascicolo: 3, volume: 266, anno: 2001,
pagine: 425 - 435
SICI:
1617-4615(200111)266:3<425:PCCFOR>2.0.ZU;2-S
Fonte:
ISI
Lingua:
ENG
Soggetto:
CAMPESTRIS PV CAMPESTRIS; GRAM-NEGATIVE BACTERIA; ESCHERICHIA-COLI; XANTHOMONAS-CAMPESTRIS; TERMINUS REGION; NUCLEOTIDE-SEQUENCE; ADSORPTION PROTEIN; BINDING PROTEIN; COAT PROTEIN; GENE;
Keywords:
Xanthomonas; filamentous phage; Xer-specific integration system; homologous recombination;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
53
Recensione:
Indirizzi per estratti:
Indirizzo: Tseng, YH Natl Chunghsing Univ, Inst Mol Biol, Taichung 402, Taiwan Natl Chunghsing Univ Taichung Taiwan 402 Taichung 402, Taiwan
Citazione:
N.T. Lin et al., "Plasmids carrying cloned fragments of RF DNA from the filamentous phage phi Lf can be integrated into the host chromosome via site-specific integration and homologous recombination", MOL GENET G, 266(3), 2001, pp. 425-435

Abstract

Different regions of RF DNA from the filamentous bacteriophage phi Lf werecloned in Escherichia coli vectors that can not be maintained in Xanthomonas. After introduction into X. campestris pv. campestris 17 (Xc17), most ofthese constructs were found to integrate into the host chromosome, either by recA-dependent homologous recombination or recA-independent site-specific integration, Mutations in himA, which codes for the alpha -subunit of theIntegration Host Factor, does not affect the integration, Integration occurs into a chromosomal region which harbors a copy of a defective phage (4445 bp) that shares a high degree of identity with the phi Lf genome. While various parts of the 4445-bp region are susceptible to homologous recombination, site-specific integration requires the attB sequence on the chromosomeand the phage attP. The attB shows a high level of sequence identity (22 out of 28 bp) to the dif site required for E. coli Xer site-specific recombination, including the 6-bp central region, and 8/11 identity in both the left XerC-binding arm and the right XerD-binding arm, with the innermost 5 ntof the arms forming a dyad symmetry that is also present in dif. The attP has the same central region and shows 10/11 identity to the dif site in theleft arm, but the sequence of the right arm is less conserved than that ofattB. The smallest regions still capable of mediating integration are a cloned 72-bp phi Lf attP-containing sequence and a 51-bp Xc17 attB-containingsequence, which was reinserted into the Xc17 chromosome after the 4445-bp region had been deleted, indicating that accessory sequences are not necessary and that the integrase required for site-specific integration is neither specified by the 4445-bp Xc17 chromosomal region nor encoded by the phi Lf genome.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 08/04/20 alle ore 07:19:36