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KMID : 0357319940290060591
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Journal of the Korean Society for Microbiology
1994 Volume.29 No. 6 p.591 ~ p.602
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Nucleotide Sequence Analysis and Expression of 56 kDa Antigen Gene of Rickettsia tsutsugamushi Strain TA716 in Escherichia coli
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엄기혁
장원종/성승용/박세광/차창룡/장우현/최명식
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Abstract
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The gene encoding the 56 kilodalton(kDa) antigen of Rickettsia tsutsugamushi strain TA716 was cloned, sequenced and expressed in Escherichia coli in the form of maltose binding protein (MBP) fusion protein.
The 56kDa antigen gene of R. tsutsugamushi strain TA716 was amplified by polymerase chain reaction(PCR) with the primers that were synthesized on the basis of the sequence of strain Gilliam, Karp, and Boryong 56kDa antigen gene. These amplified
DNA
fragments were cloned into pBluescript vector for sequence analysis and were designated as pBT716G and were cloned into pIH821 plasmid vector for expression and was designated as pMT716C.
The size of open reading frame of the 56kDa antigen gene of strain TA716 was 1,575bp and the protein was composed of 524 amino acids. The deduced molecular weight of the 56kDa antigen of strain TA716 was 55,960. There was a leader sequence which
consisted of 22 amino acids.
Alternate hydrophobic and hydrophilic regions were observed and we could presume that the 56kDa protein is a transmembrane protein.
The nucleotide sequence and amino acid sequence were compared with that of strain Gilliam, Karp, Kato. Kawasaki, and Boryong. The nucleotide sequence homology varied 78.5-87.5% and the deduced amino acid sequence homology varied 68.0-80.8%.
The 56kDa protein of R. tsutsugamushi strain TA716 expressed in E. coli in the from of MBP fusion protein reacted with 716-D, a specific monoclonal antibody to R. tsutsugamush strain TA716.
The basic informations on R. tsutsugamushi strain TA716 obtained from this study will be useful in the development of diagnostic tools and vaccine for scrub typhus.
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