Submit Manuscript

Research Journal of Microbiology

Year: 2015 | Volume: 10 | Issue: 11 | Page No.: 551-556
Short Communication

Direct Cloning Protocol for Rapid Transformant Selection

Ahmed E. Gomaa
Department of Biotechnology, Chonnam National University, Chonnam, 550-749, Republic of Korea
Seung Hwan Yang
Center for Nutraceutical and Pharmaceutical Materials, Myongji University, Gyeonggi, 449-728, Republic of Korea
Sang Mi Sun
Department of Biotechnology, Chonnam National University, Chonnam, 550-749, Republic of Korea
Gyuhwa Chung
Department of Biotechnology, Chonnam National University, Chonnam, 550-749, Republic of Korea
This study highlights a reporter-less direct-cloning protocol based on PCR and digestion/ligation reactions. Few manipulations were done allowing the user to reduce the time required for screening the right colony of any transformed cells. The obtained result showed high efficiency compared to the other commercially-available high throughput techniques, especially Gateway.
PDF Fulltext XML References Citation

How to cite this article

Ahmed E. Gomaa, Seung Hwan Yang, Sang Mi Sun and Gyuhwa Chung, 2015. Direct Cloning Protocol for Rapid Transformant Selection. Research Journal of Microbiology, 10: 551-556.

URL: https://scialert.net/abstract/?doi=jm.2015.551.556

Leave a Comment

Your email address will not be published. Required fields are marked *

Article Trend

Total views 3159

References

  1. Balagurumoorthy, P., S.J. Adelstein and A.I. Kassis, 2008. Method to eliminate linear DNA from mixture containing nicked circular, supercoiled and linear plasmid DNA. Anal. Biochem., 381: 172-174.
    CrossRefDirect Link
  2. Costa, G.L., A. Grafsky and M.P. Weiner, 1994. Cloning and analysis of PCR-generated DNA fragments. Genome Res., 3: 338-345.
    Direct Link
  3. Datsenko, K.A. and B.L. Wanner, 2000. One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc. Natl. Acad. Sci. USA., 97: 6640-6645.
    CrossRefPubMedDirect Link
  4. Delorme, E., 1989. Transformation of Saccharomyces cerevisiae by electroporation. Applied Environ. Microbiol., 55: 2242-2246.
    Direct Link
  5. Demple, B. and L. Harrison, 1994. Repair of oxidative damage to DNA: Enzymology and biology. Ann. Rev. Biochem., 63: 915-948.
    CrossRefPubMedDirect Link
  6. Esposito, D., L.A. Garvey and C.S. Chakiath, 2009. Gateway cloning for protein expression. Methods Mol. Biol., 498: 31-54.
    CrossRefDirect Link
  7. Gunn, L., J. Whelden and J.A. Nickoloff, 1995. Transfer of episomal and integrated plasmids from Saccharomyces cerevisiae to Escherichia coli by electroporation. Methods Mol. Biol., 47: 55-66.
    CrossRefDirect Link
  8. Ito, T., T. Chiba, R. Ozawa, M. Yoshida, M. Hattori and Y. Sakaki, 2001. A comprehensive two-hybrid analysis to explore the yeast protein interactome. Proc. Natl. Acad. Sci., 98: 4569-4574.
    CrossRefDirect Link
  9. Mirhendi, H., K. Diba, A. Rezaei, N. Jalalizand, L. Hosseinpur and H. Khodadadi, 2007. Colony PCR is a rapid and sensitive method for DNA amplification in yeasts. Iran. J. Public Health, 36: 40-44.
  10. Liu, P., N.A. Jenkins and N.G. Copeland, 2003. A highly efficient recombineering-based method for generating conditional knockout mutations. Genome Res., 13: 476-484.
    Direct Link
  11. Liu, Q., M.Z. Li, D. Leibham, D. Cortez and S.J. Elledge, 1998. The univector plasmid-fusion system, a method for rapid construction of recombinant DNA without restriction enzymes. Curr. Biol., 8: 1300-1309.
    CrossRefDirect Link
  12. Liu, Q., M.Z. Li, D. Liu and S.J. Elledge, 2000. Rapid construction of recombinant DNA by the univector plasmid-fusion system. Methods Enzymol., 328: 530-549.
    CrossRefPubMedDirect Link
  13. Luo, G. and T.G. Mitchell, 2002. Rapid identification of pathogenic fungi directly from cultures by using multiplex PCR. J. Clin. Microbiol., 40: 2860-2865.
    CrossRefDirect Link
  14. Marsischky, G. and J. LaBaer, 2004. Many paths to many clones: A comparative look at high-throughput cloning methods. Genome Res., 14: 2020-2028.
    Direct Link
  15. Martzen, M.R., S.M. McCraith, S.L. Spinelli, F.M. Torres, S. Fields, E.J. Grayhack and E.M. Phizicky, 1999. A biochemical genomics approach for identifying genes by the activity of their products. Science, 286: 1153-1155.
    CrossRefDirect Link
  16. Mol, C.D., C.F. Kuo, M.M. Thayer, R.P. Cunningham and J.A. Tainer, 1994. Structure and function of the multifunctional DNA-repair enzyme exonuclease III. Nature, 374: 381-386.
    CrossRefDirect Link
  17. Osek, J., 2001. Multiplex polymerase chain reaction assay for identification of enterotoxigenic Escherichia coli strains. J. Vet. Diagn. Invest., 13: 308-311.
    CrossRefDirect Link
  18. Reboul, J., P. Vaglio, N. Tzellas, N. Thierry-Mieg and T. Moore et al., 2001. Open-reading-frame sequence tags (OSTs) support the existence of at least 17,300 genes in C. elegans. Nat. Genet., 27: 332-336.
    CrossRefDirect Link
  19. Uetz, P., L. Giot, G. Cagney, T.A. Mansfield and R.S. Judson et al., 2000. A comprehensive analysis of protein-protein interactions in saccharomyces cerevisiae. Nature, 403: 623-627.
    CrossRefDirect Link
  20. Upcroft, P. and A. Healey, 1987. Rapid and efficient method for cloning of blunt-ended DNA fragments. Gene, 51: 69-75.
    CrossRefDirect Link
  21. Walhout, A.J.M., G.F. Temple, M.A. Brasch, J.L. Hartley, M.A. Lorson, S. van den Heuvel and M. Vidal, 2000. GATEWAY recombinational cloning: Application to the cloning of large numbers of open reading frames or ORFeomes. Methods Enzymol., 328: 575-592.
    CrossRefDirect Link
  22. You, C., X.Z. Zhang and Y.H.P. Zhang, 2012. Simple cloning via direct transformation of PCR product (DNA Multimer) to Escherichia coli and Bacillus subtilis. Applied Environ. Microbiol., 78: 1593-1595.
    CrossRefDirect Link
  23. Zhang, Y., J.P. Muyrers, G. Testa and A.F. Stewart, 2000. DNA cloning by homologous recombination in Escherichia coli. Nat. Biotechnol., 18: 1314-1317.
    CrossRefDirect Link
  24. Zhu, H., M. Bilgin, R. Bangham, D. Hall and A. Casamayor et al., 2001. Global analysis of protein activities using proteome chips. Science, 293: 2101-2105.
    CrossRefDirect Link

Keywords