Abstract
The DNA increment method, designed for measuring the increment in the amount of DNA after inhibition of initiation of fresh rounds of replication initiation was employed to measure the rate of deoxyribonucleic acid (DNA) chain growth in Mycobacterium tuberculosis H37Rv growing in Youman and Karlson's medium at 37°C with a generation time of 24 h and also in relatively fast growing species like Mycobacterium smegmatis and Escherichia coli. From the results obtained, the time required for a DNA replication fork to traverse the chromosome from origin to terminus (C period) was calculated. The chain elongation rates of DNA of the three organisms was determined from the C period and the known genome sizes assuming that all these genomes have a single replication origin and bidirectional replication fork. The rate for M. tuberculosis was 3,200 nucleotides per min about 11 times slower than that of M. smegmatis and about 13–18 times slower than that of E. coli.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
Abbreviations
- DNA:
-
deoxyribonucleic acid
- td:
-
delay in initiation
- OD:
-
optical density
- CAM:
-
chloramphenicol
- RIF:
-
rifampicin
References
Bagdasarian MM, Izakowska M, Bagdasarian M (1977) Suppression of the DNA A phenotype by mutations in the rpoB cistron of ribonucleic acid polymerase in Salmonella typhimurium and Escherichia coli. J Bacteriol 130:577–582
Barksdale L, Kim KS (1977) Mycobacteria. Bacteriol Rev 41:217–372
Bremer H, Churchward G (1977) Deoxyribonucleic acid synthesis after inhibition of rounds of replication in Escherichia coli. J Bacteriol 130:692–697
Churchward G, Bremer H (1977) Determination of deoxyribonucleic acid replication time in exponentially growing Escherichia coli B/r. J Bacteriol 130:1206–1213
Gradwohl RBH (1948) Clinical laboratory methods and diagnosis, 4th edn, vol II. C. V. Mosby Company. St. Louis, MO, p 1356
Harshey RM, Ramakrishnan T (1977) Rate of ribonucleic acid chain growth in Mycobacterium tuberculosis H37Rv. J Bacteriol 129:616–622
Kornberg A (1980) DNA replication. Freeman and Co., San Francisco, CA, p 446
Lane HED, Dernhardt DT (1975) The rep. mutation. IV. Slower movement of replication forks in Escherichia coli rep. strains. J Mol Biol 97:99–112
Maaløe O, Hanawalt PC (1961) Thymine deficiency and the normal DNA replication cycle. J Mol Biol 3:144–155
Pritchard RH, Zaritsky A (1971) Effect of thymine concentration on the replication velocity of DNA in a thymineless mutant of Escherichia coli. Nature (Lond) 226:126–131
Projan SJ, Wechler JA (1981) RNA polymerase is required for DNA initiation in vitro. Mol Gen Gent 183:78–81
Seiki M, Ogasawara N, Yoshikawa H (1981) Structure and function of the region of the replication orgin of the Bacillus subtilis chromosome. Mol Gen Gent 183:227–233
Tamonoi F, Saito H, Richardson CC (1980) Physical mapping of primary and secondary origins of bacteriophage T7 DNA replications. Proc Natl Acad Sci USA 77:2656–2660
Woodley CL, Baldwin JN, Greenberg J (1981) Nitrosoguanidine sequential mutagenesis mapping of Mycobacterium tuberculosis genes. J Bacteriol 147:176–180
Youmans GP, Karlson AG (1947) Streptomycin sensitivity of tubercle bacilli. Studies on recently isolated tubercle bacilli in vivo. Amer Rev Tub Pulm Dis 55:529–533
Youmans GP, Youmans AS (1949) A method for the determination of the rate of growth of tubercle bacilli by the use of small inocula. J Bacteriol 58:247–255
Zaritsky A, Pritchard RH (1971) Replication time of the chromosome in thymineless mutants of Escherichia coli. J Mol Biol 60:65–74
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hiriyanna, K.T., Ramakrishnan, T. Deoxyribonucleic acid replication time in Mycobacterium tuberculosis H37 Rv. Arch. Microbiol. 144, 105–109 (1986). https://doi.org/10.1007/BF00414718
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00414718