Reports of Biochemistry and Molecular Biology
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Basic Sciences
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admin
2322-3480
2322-3480
10.61186/rbmb
en
jalali
1398
2
1
gregorian
2019
5
1
8
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online
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fulltext
en
Design and Construction of a Eukaryotic Cloning Vector Encoding the mpt51 Gene of <em>Mycobacterium tuberculosis</em>
میکروب شناسی
Microbiology
مقالات اصلی
Original Article
<div style="text-align: justify;"><strong><em>Background:</em></strong> Tuberculosis (TB) is the leading cause of death by infectious diseases worldwide, and especially prevalent in developing countries. Several vaccines against TB have been developed, recently. The aim of the present study was to design and construct a cloning vector encoding <em>Mycobacterium tuberculosis </em>(MTB)<em> mpt51 </em>gene.<br>
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<strong><em>Methods:</em></strong> DNA was extracted from MTB H37Rv strain. Gene-specific primers were designed using Gene Runner software and the <em>mpt51 </em>gene was amplified by PCR. The amplified fragment and pcDNA3.1(+) cloning vector were both digested with restriction enzymes, the <em>mpt51 </em>fragment was ligated into the vector, and the <em>Escherichia coli</em> (<em>E. coli) </em>TOP10 strain were transformed by the recombinant plasmid. Positive clones were identified by colony PCR, restriction enzyme digestion, and DNA sequencing.<br>
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<strong><em>Results:</em></strong> The<em> mpt51</em> gene was successfully cloned into pcDNA3.1(+). A 6400 bp band for the pcDNA3.1(+)/<em>mpt51 </em>recombinant plasmid and a 926 bp band for <em>mpt51 </em>were observed by colony PCR, and restriction enzyme digestion on agarose gels. The DNA sequence was 100% homologous with the <em>mpt51</em> fragment of H37Rv in GenBank.<br>
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<strong><em>Conclusions:</em></strong> In the current study, the <em>mpt51 </em>gene of MTB was correctly cloned into pcDNA3.1(+). The expression of this recombinant vector can be studied in eukaryotic cells. Moreover, it is possible to determine the efficacy of this vector as a DNA vaccine candidate, and to test its protective function compared to BCG in animal models in future.<br>
</div>
Molecular Cloning, MPT51 antigen, Mpt51 gene, Mycobacterium tuberculosis.
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http://rbmb.net/browse.php?a_code=A-10-69-1&slc_lang=en&sid=1
Faria
Hasanzadeh Haghighi
HasanzadehF1@mums.ac.ir
10031947532846006535
10031947532846006535
No
Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
Ehsan
Aryan
Aryane@mums.ac.ir
10031947532846006536
10031947532846006536
No
Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
Aida
Gholoobi
GhasemiF891@mums.ac.ir
10031947532846006537
10031947532846006537
No
Department of Modern Sciences and Technologies, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
Hosna
Zare
ZareH951@mums.ac.ir
10031947532846006538
10031947532846006538
No
Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
Zahra
Meshkat
meshkatz@mums.ac.ir
10031947532846006539
10031947532846006539
Yes
Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran and Department of Microbiology and Virology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.