TY - JOUR
T1 - 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
AU - Santhosh, Ramachandran Sarojini
AU - Pandian, Shunmugiah Karutha
AU - Lini, Nirmala
AU - Shabaana, Abdul Khader
AU - Nagavardhini, Avuthu
AU - Dharmalingam, Kuppamuthu
N1 - Funding Information:
R.S.S. acknowledges the Council of Scientific and Industrial Research, New Delhi, India for the award of a Senior Research Fellowship. N.L. thanks University Grants Commission, New Delhi for the award of Junior Research Fellowship. This research was funded by a Centre for Genetic Engineering and Strain Manipulation grant from the Department of Biotechnology, Government of India, New Delhi, India to K. Dharmalingam (BT/03/002/87-Vol. IV). Grant from Heiser program for Research in Leprosy and Tuberculosis, New York, USA is acknowledged.
PY - 2005/8/1
Y1 - 2005/8/1
N2 - Plasmid pSET152 is a broad host range mobilizable vector which integrates into streptomyces chromosome utilizing att site and int function of ØC31. 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.
AB - Plasmid pSET152 is a broad host range mobilizable vector which integrates into streptomyces chromosome utilizing att site and int function of ØC31. 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.
KW - Integrase
KW - Mammalian cell entry
KW - Mycobacterium leprae
KW - Mycobacterium smegmatis
KW - RT-PCR
KW - pSET152
UR - http://www.scopus.com/inward/record.url?scp=22844437189&partnerID=8YFLogxK
U2 - 10.1016/j.femsim.2005.05.004
DO - 10.1016/j.femsim.2005.05.004
M3 - Article
C2 - 15949925
AN - SCOPUS:22844437189
SN - 0928-8244
VL - 45
SP - 291
EP - 302
JO - FEMS Immunology and Medical Microbiology
JF - FEMS Immunology and Medical Microbiology
IS - 2
ER -