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Penelitian

Purkan, S.Si, M.Si

Fakultas Fakultas Sains dan Teknologi / Departemen Kimia

Tim Peneliti :

  1. Purkan
  2. Wiwin Retnowati

Tahun : 2009

Halaman Naskah : 40 halaman

Sumber Dana : DIPA-RM STRATNAS

Besaran Dana : 0

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Kategori Penelitian : Kesehatan

Posting : 02-12-2010

Visitor : 5129


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Title :

Mechanism of Isoniazid Resistance in Mycobacterium tuberculosis: analysis of katG gene mutation and its Bearing With Biochemical Properties Mutant Enzymes of Recombinant Catalase-Peroxidase

Author : Purkan, S.Si, M.Si


Year : 2009

Abstact :

Tuberculosis is an infectious disease which caused by Mycobacterium tuberculosis and kills many people in the world. Drug resistant of MM tuberculosis is serious problem on TB therapy. Multi drug resistant (MDR) TB mean as M. tuberculosis which has resistant phenotype toward two first line drugs, rifampin and isoniazid. The NOR phenotype of the strains comes from accumulation of mutation of anti TB drug-targeted genes. Some clinical isolates of M. tuberculosis have isoniazid resistant phenotype, but their causative have unknown yet. The aims of the research are to know the causative and mechanism of isoniazid resistance to clinical isolates by determining of katG mutation on genotype level and characterizing of mutant catalase-peroxidase activity on protein level, moreover to know the correlation between katG mutation profile as well as catalase-peroxidase activity with isoniazid resistance. In the first year, the work is oriented to identify mutation of katG gene belong to clinical isolates and analyze the correlation of katG mutation with isoniazid resistance through in silico analysis. The work steps for the research in the first year consist of amplifying of katG genes, cloning of katG gene on pGemT vector, restriction analyzing of recombinant DNA pGemT-katG, katG sequencing, mutation analyzing, subcloning of katG gen in the pET20b vector, and resequencing of katG in the last work. The katG gene of isoniazid resistant M. tuberculosis clinical isolates (L8, L19, L 10, L21, R2 and R9) have 2.2 kb in size, had amplified well by PCR technique using a pair of primer FG and RG. Molecular characterization of katG gene through cloning and subcloning showed the variant on katG nucleotides of clinical isolates. Nucleotide mutations which found on katG gene of clinical isolate are assumed as prime causative of the isoniazid resistance appearance. In silico translation yield toward katG nucleotides sequence of clinical isolates showed the amino acid mutations in N domain of KatG protein are inclined increasing Isoniazid resistance comparing to mutation in C domain. Mutation which change amino acids near to critical residues of KatG protein is also inclined increasing Isoniazid resistance comparing to mutation in far region of critical sides. The mutations located in katG gene, which alter amino acid residues in both N and C domains of protein KatG leading to increase levels of isoniazid resistance in ll~f. tuberculosis comparing to mutation in one of the domains. The increasing of the number of mutation in katG gene is inclined enhancing of Isoniazid resistance. Further research which explore the aspect of KatG protein activity related to expression, pufication and characterization of KatG protein is needed to obtain the comprehensive mechanism of Isoniazid resistance.


Keyword : Tuberculosis is an infectious, genotype level ,


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