Cloning of mce1 locus of Mycobacterium leprae in Mycobacterium smegmatis mc2 155 SMR5 and evaluation of expression of mce1 genes in M. smegmatis and M. leprae
{"title":"Cloning of mce1 locus of Mycobacterium leprae in Mycobacterium smegmatis mc2 155 SMR5 and evaluation of expression of mce1 genes in M. smegmatis and M. leprae","authors":"Ramachandran Sarojini Santhosh, Shunmugiah Karutha Pandian , Nirmala Lini, Abdul Khader Shabaana , Avuthu Nagavardhini, Kuppamuthu Dharmalingam","doi":"10.1016/j.femsim.2005.05.004","DOIUrl":null,"url":null,"abstract":"<div><p><span>Plasmid pSET152 is a broad host range mobilizable vector which integrates into streptomyces chromosome utilizing </span><em>att</em> site and <em>int</em> function of ∅C31. Transformation of this plasmid into <span><em>Mycobacterium </em><em>smegmatis</em></span> mc<sup>2</sup> 155 SMR5 gave stable transformants carrying the pSET152 as an integrated copy. Integration occurred at the cross over sequence 5′TTG disrupting the <em>gatA</em> gene (Glu-tRNA<sup>Gln</sup> amidotransferase subunitA), which is non-essential under conditions used. Recombinant pSET152 plasmids carrying <em>mce1</em> locus of <span><em>Mycobacterium leprae</em></span> were used to construct <em>M. smegmatis</em> transformants carrying the <em>mce1</em> locus in their chromosome. RT-PCR analysis revealed specific transcripts of <em>M. leprae mce</em> in <em>M. smegmatis</em><span>. The transcribed mRNA carried intergenic regions between genes of </span><em>mce1</em> locus indicating that <em>mce1</em> locus is an operon. Examination of <em>M. leprae</em><span> specific mRNA from lepromatous leprosy patient’s biopsy showed that </span><em>mce</em> locus is transcribed as an operon in the pathogen also.</p></div>","PeriodicalId":12220,"journal":{"name":"FEMS immunology and medical microbiology","volume":"45 2","pages":"Pages 291-302"},"PeriodicalIF":0.0000,"publicationDate":"2005-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.femsim.2005.05.004","citationCount":"9","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"FEMS immunology and medical microbiology","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0928824405001203","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 9
Abstract
Plasmid pSET152 is a broad host range mobilizable vector which integrates into streptomyces chromosome utilizing att site and int function of ∅C31. Transformation of this plasmid into Mycobacterium smegmatis mc2 155 SMR5 gave stable transformants carrying the pSET152 as an integrated copy. Integration occurred at the cross over sequence 5′TTG disrupting the gatA gene (Glu-tRNAGln amidotransferase subunitA), which is non-essential under conditions used. Recombinant pSET152 plasmids carrying mce1 locus of Mycobacterium leprae were used to construct M. smegmatis transformants carrying the mce1 locus in their chromosome. RT-PCR analysis revealed specific transcripts of M. leprae mce in M. smegmatis. The transcribed mRNA carried intergenic regions between genes of mce1 locus indicating that mce1 locus is an operon. Examination of M. leprae specific mRNA from lepromatous leprosy patient’s biopsy showed that mce locus is transcribed as an operon in the pathogen also.
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