Cloning and sequence analysis of the culture filter protein 10 gene from Mycobacterium tuberculosis H37Ra

Objective: To obtain the culture filter protein 10( CFP10) gene from Mycobacterium tuberculosis H37Ra and clone it into plasmid for nucleotide sequence analysis. Methods: The genomic DNA from Mycobacterium tuberculosis H37Ra was extracted as template,and the primer was designed according to the CFP10 gene of Mycobacterium tuberculosis H37Ra. The CFP10 gene was amplified from the genome of Mycobacterium tuberculosis H37Ra by polymerase chain reaction( PCR) and cloned into pTG19-T vector. After identification by PCR and restriction enzyme analysis,the CFP10 gene was sequenced,analyzed and compared with that of H37Ra reported in GenBank. Results: The sequencing result showed that the gene was obtained with an open reading frame of 303 bp. Compared with the DNA sequence of Mycobacterium tuberculosis H37Ra,the homology was 100%,which deduced that the amino acid sequence of CFP10 was 100% identity. Conclusions: The CFP10 gene of Mycobacterium tuberculosis H37Ra had been cloned successfully and the homology was 100% compared with Mycobacterium tuberculosis H37Ra.