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Study of carD gene sequence in clinical isolates of Mycobacterium tuberculosis
- Source :
- Acta microbiologica et immunologica Hungarica. 61(1)
- Publication Year :
- 2014
-
Abstract
- Mycobacterium tuberculosis growth rate is closely coupled to rRNA transcription which is regulated through carD gene. The aim of this study was to determine the sequence of carD gene in drug susceptible and resistant clinical isolates of M. tuberculosis and designing of a PCR assay based on carD sequence for rapid detection of this bacterium.Specific primers for amplification of carD gene were carefully designed, so that whole sequence of gene could be amplified; therefore primers were positioned at the upstream (promoter of this gene and ispD gene) and downstream (in ispD gene). DNA from 41 clinical isolates of M. tuberculosis with different pattern of drug resistance was used in the study. PCR conditions and annealing temperature were designed by means of online programs. PCR products were sequenced by ABI system.PCR product of carD gene was a 524 bp fragment. This method could detect all resistant and susceptible strains of M. tuberculosis. The size of amplified fragment was similar in all investigated samples. Sequence analysis showed that there was similar sequence in all of our isolates therefore probably this gene is considered to be conservative. Translation of nucleotide mode to amino acids was showed that TRCF domain in N-terminal of protein CarD was found to be fully conservative.This is the first study on the sequence of carD gene in clinical isolates of M. tuberculosis. This conservative gene is recommended for use as a target for designing of suitable inhibitors as anti-tuberculosis drug because its importance for life of MTB. In the other hand, a PCR detection method based on detection of carD gene was recommended for rapid detection in routine test.
- Subjects :
- Genetics
Tuberculosis
General Immunology and Microbiology
biology
Base Sequence
Pcr cloning
Molecular Sequence Data
General Medicine
Drug resistance
Mycobacterium tuberculosis
Sequence Analysis, DNA
medicine.disease
biology.organism_classification
RRNA transcription
chemistry.chemical_compound
chemistry
Bacterial Proteins
Genes, Bacterial
medicine
Humans
Amino Acid Sequence
Gene sequence
Gene
DNA
Subjects
Details
- ISSN :
- 12178950
- Volume :
- 61
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Acta microbiologica et immunologica Hungarica
- Accession number :
- edsair.doi.dedup.....c245a2d2ec2d8cbf7dc3bdd93a493748