%0 Journal Article %A Alexander, Dylan C. %A Devlin, David J. %A Hewitt, Duane D. %A Horan, Ann C. %A Hosted, Thomas J. %T Development of the Micromonospora carbonacea var. africana ATCC 39149 bacteriophage pMLP1 integrase for site-specific integration in Micromonospora spp. %D 2003 %J Microbiology, %V 149 %N 9 %P 2443-2453 %@ 1465-2080 %R https://doi.org/10.1099/mic.0.26318-0 %K apr, apramycin resistance gene %K attB, bacterial attachment site %K oriT, origin of transfer %K attP, phage attachment site %K ApR, apramycin resistance %K att/int, region containing the excisionase, integrase gene and attP site %K HmR, hygromycin resistance %K MCS, multiple cloning site %K hyg, hygromycin resistance gene %K PKS, polyketide synthase %K AmR, ampicillin resistance %I Microbiology Society, %X Micromonospora carbonacea var. africana 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 KpnI fragment revealed pMLP1 att/int functions consisting of an integrase, an excisionase and the phage attachment site (attP). Plasmids pSPRH840 and pSPRH910, containing the pMLP1 att/int region, were introduced into Micromonospora spp. by conjugation from Escherichia coli. Sequence analysis of DNA flanking the integration site confirmed site-specific integration into a tRNAHis gene in the chromosome. The pMLP1 attP element and chromosomal bacterial attachment (attB) site contain a 24 bp region of sequence identity located at the 3′ end of the tRNA. Integration of pMLP1-based plasmids in M. carbonacea var. africana caused a loss of the pMLP1 phage. Placement of an additional attB site into the chromosome allowed integration of pSPRH840 into the alternate attB site. Plasmids containing the site-specific att/int functions of pMLP1 can be used to integrate genes into the chromosome. %U https://www.microbiologyresearch.org/content/journal/micro/10.1099/mic.0.26318-0