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Volume 149, Issue 9

Development of the var. ATCC 39149 bacteriophage pMLP1 integrase for site-specific integration in spp. Free

Abstract

var. ATCC 39149 contains a temperate bacteriophage, pMLP1, that is present both as a replicative element and integrated into the chromosome. Sequence analysis of a 4·4 kb I fragment revealed pMLP1 / functions consisting of an integrase, an excisionase and the phage attachment site (). Plasmids pSPRH840 and pSPRH910, containing the pMLP1 / region, were introduced into spp. by conjugation from . Sequence analysis of DNA flanking the integration site confirmed site-specific integration into a tRNA gene in the chromosome. The pMLP1 element and chromosomal bacterial attachment () site contain a 24 bp region of sequence identity located at the 3′ end of the tRNA. Integration of pMLP1-based plasmids in var. caused a loss of the pMLP1 phage. Placement of an additional site into the chromosome allowed integration of pSPRH840 into the alternate site. Plasmids containing the site-specific / functions of pMLP1 can be used to integrate genes into the chromosome.

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  • Accepted:
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  • Published Online:
Keyword(s): AmR, ampicillin resistance , ApR, apramycin resistance , apr, apramycin resistance gene , att/int, region containing the excisionase, integrase gene and attP site , attB, bacterial attachment site , attP, phage attachment site , HmR, hygromycin resistance , hyg, hygromycin resistance gene , MCS, multiple cloning site , oriT, origin of transfer and PKS, polyketide synthase
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2003-09-01
2024-04-24
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Development of the Micromonospora carbonacea var. africana ATCC 39149 bacteriophage pMLP1 integrase for site-specific integration in Micromonospora spp.
Microbiology 149, 2443 (2003); https://doi.org/10.1099/mic.0.26318-0
/content/journal/micro/10.1099/mic.0.26318-0
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