Journal List > Tuberc Respir Dis > v.60(2) > 1000908

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Lee, Kim, Park, and Bai: Investigation of the Growth Rate Change in Recombinant BCG which was cloned Mycobacterium tuberculosis Adenylate Kinase Mutation Gene or Human Muscle-type Adenylate Kinase Synthetic Gene
Original Article
Tuberculosis and Respiratory Diseases

Tuberculosis and Respiratory Diseases 2006; 60(2): 187-193.

Published online: 28 February 2006

DOI: https://doi.org/10.4046/trd.2006.60.2.187

Investigation of the Growth Rate Change in Recombinant BCG which was cloned Mycobacterium tuberculosis Adenylate Kinase Mutation Gene or Human Muscle-type Adenylate Kinase Synthetic Gene

Seung-Heon Lee1, Hyo-Joon Kim2, Young-Kil Park1, Gill-Han Bai1

1Department of Molecular Biology, Korean Institute of Tuberculosis, Seoul, Korea.

2Department of Biochemistry, Hanyang University College of Biotechnology, Ansan, Korea.

Corresponding author: Gill-Han Bai. Address: Korean Institute of Tuberculosis 14 Woomyundong, Sochogu, Seoul, 137-140, Korea. Phone: +82-2-577-5766, Fax: +82-2-573-1914, gbai@hotmail.com

Received 2 November 2005       Accepted 27 January 2006

Copyright © 2006 The Korean Academy of Tuberculosis and Respiratory Diseases

Abstract

Background

Normal cell proliferation and viability is strongly depends on the availability of metabolic energy and the maintenance of the appropriate adenylate-nucleotide pools. Hypothetically, changes in adenylate kinase (AK) expression could therefore be associated with adaptation to altered growth characteristics or inversely altered growth characteristics of proliferating cells could drive the changes in the metabolic profile. This study investigated whether the expression of either AK1 or a Mycobacterium tuberculosis adenylate kinase mutant which has the same catalytic activity of AK1 could affect the growth rate of slow-growing BCG.

Method

Recombinant BCGs, which were cloned the human muscle-type adenylate kinase synthetic gene (AK1) and adenylate kinase mutation gene (AKmtDM) of Mycobacterium tuberculosis into the Mycobacterium/E.coli expression vectors, were constructed. Recombinant BCGs and wild-type BCG were cultured in 7H9 media and the optical density at 600nm was measured at intervals of 2-3 days.

Result

There wasn't the growth rate change induced by AK1 or AKmtDM expression in recombinant BCGs.

Conclusion

The expression of AK1 or Mycobacterium tuberculosis adenylate kinase mutant in BCG does not affect the growth rate of BCG.

Keywords: Adenylate kinase, Mutation, Growth rate, BCG

Figures and Tables

Figure 1
Construction of recombinant plasmids.
KanR : Tn903 derived aph gene conferring kanamycin resistance as a selectable marker.
OriM : The origin of mycobacterial replication derived from pAL5000.
Phsp60 : Mycobacterial promoter derived from regulatory sequences of a BCG heat shock protein, hsp60.
MCS : Multiple cloning sites.
trd-60-187-g001
Figure 2
Growth curve of recombinant BCGs and values of optical density at 600nm. Data represent the mean of triplicate values ± standard error of the mean.
trd-60-187-g002
Table 1
Primers for cloning
trd-60-187-i001

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