Share This Article:

Construction of a Shuttle Vector for Heterologous Gene Expression in Escherichia coli and Microalgae Anabaena

Full-Text HTML Download Download as PDF (Size:639KB) PP. 540-544
DOI: 10.4236/eng.2013.510B111    4,218 Downloads   5,354 Views   Citations

ABSTRACT

Theconstruction of an integrative shuttle expression vector and potential utility was reported inEscherichiacoliandAnabaena(Nostoc) sp. strain PCC 7120. The vector comprised of the following elements: (a) an intergenic non-coding region fromAnabaenato facilitate its genomic integration (b) a strong functional PpsbAIpromoter fromAnabaenafor desired gene expression and (c) neomycin phosphotransferase gene with its own promoter for the selection of transfor-mants. The constructed vectorpAnFP was evaluated by cloning, transfer and expression of thegfpgene encoding green fluorescent protein. When theE.coliandAnabaenasp. strain PCC 7120 were transformed, intensive green fluorescence produced by the products of GFP protein was observed. This result indicated that the integrative shuttle vector pAnFP can be promisingly used in genome transformation for expression of heterologous genes inE.coliand microalgae such asAnabaenaandNostocstrains.

Conflicts of Interest

The authors declare no conflicts of interest.

Cite this paper

Song, D. , Li, J. , Hu, X. and Xi, B. (2013) Construction of a Shuttle Vector for Heterologous Gene Expression in Escherichia coli and Microalgae Anabaena. Engineering, 5, 540-544. doi: 10.4236/eng.2013.510B111.

References

[1] P. Bhadury and P. C. Wright, “Exploitation of Marine Algae: Biogenic Compounds for Potential Antifouling Applications,” Planta, Vol. 219, No. 4, 2004, pp. 561-578. http://dx.doi.org/10.1007/s00425-004-1307-5
[2] A. Singh, P. S. Nigam and J. D. Murphy, “Renewable Fuels from Algae: An Answer to Debatable Land Based Fuels,” Bioresource Technology, Vol. 102, No. 1, 2011, pp. 10-16. http://dx.doi.org/10.1016/j.biortech.2010.06.032
[3] D. H. Song, J. J. Fu and D. J. Shi, “Exploitation of Oil- Bearing Microalgae for Biodiesel,” Chinese Journal of Biotechnology, Vol. 24, No. 3, 2008, pp. 341-348. http://dx.doi.org/10.1016/S1872-2075(08)60016-3
[4] A. K. Chaurasia and S. K. Apte, “Improved Eco-Friendly Recombinant Anabaena PCC 7120 Strain with Enhanced Nitrogen Biofertilizer Potential,” Applied and Environmental Microbiology, Vol. 77, No. 2, 2011, pp. 395-399. http://dx.doi.org/10.1128/AEM.01714-10
[5] W. D. Stewart, “Some Aspects of Structure and Function in N2-Fixing Cyanobacteria,” Annual Review of Microbiology, Vol. 34, 1980, pp. 497-536. http://dx.doi.org/10.1146/annurev.mi.34.100180.002433
[6] P. Lindberg, P. Lindblad and L. Cournac, “Gas Exchange in the Filamentous Cyanobacterium Nostoc punctiforme Strain ATCC 29133 and Its Hydrogenase-Deficient Mutant Strain NHM5,” Applied and Environmental Microbi- ology, Vol. 70, No. 4, 2004, pp. 2137-2145. http://dx.doi.org/10.1128/AEM.70.4.2137-2145.2004
[7] H. Masukawa, K. Inoue, H. Sakurai, C. P. Wolk and R. P. Hausinger, “Site-Directed Mutagenesis of the Anabaena sp. Strain PCC 7120 Nitrogenase Active Site to Increase Photobiological Hydrogen Production,” Applied and Environmental Microbiology, Vol. 76, No. 20, 2010, pp. 6741-6750. http://dx.doi.org/10.1128/AEM.01056-10
[8] P. D. Weyman, B. Pratte and T. Thiel, “Hydrogen Production in Nitrogenase Mutants in Anabaena variabilis,” FEMS Microbiology Letters, Vol. 304, No. 1, 2010, pp. 55-61. http://dx.doi.org/10.1111/j.1574-6968.2009.01883.x
[9] A. B. M. Hossain, A. Salleh, A. N. Boyce, P. Chowdhury and M. Naqiuddin, “Biodiesel Fuel Production from Algae as Renewable Energy,” American Journal of Biochemistry and Biotechnology, Vol. 4, No. 3, 2008, pp. 250- 254. http://dx.doi.org/10.3844/ajbbsp.2008.250.254
[10] R. W. Castenholz, “Culturing of Cyanobacteria,” Methods in Enzymology, Vol. 167, 1988, pp. 68-93. http://dx.doi.org/10.1016/0076-6879(88)67006-6
[11] J. Sambrook, F. E. Fritsch and T. Maniatis, “Molecular Cloning: A Laboratory Manual,” 2nd Edition, Cold Spring Harbour Laboratory Press, New York, 1989.
[12] J. Paz-Yepes, E. Flores and A. Herrero, “Expression and Mutational Analysis of the glnB Genomic Region in the Heterocyst-Forming Cyanobacterium Anabaena sp. Strain PCC 7120, ” Journal of Bacteriology, Vol. 191, No. 7, 2009, pp. 2353-2361. http://dx.doi.org/10.1128/JB.01381-08
[13] C. C. Bauer, K. S. Ramaswamy, S. Endley, L. A. Scappino, J. W. Golden and R. Haselkorn, “Suppression of Heterocyst Differentiation in Anabaena PCC 7120 by a Cosmid Carrying Wild-Type Genes Encoding Enzymes for Fatty Acid Synthesis, ” FEMS Microbiology Letters, Vol. 151, No. 1, 1997, pp. 23-30. http://dx.doi.org/10.1111/j.1574-6968.1997.tb10390.x
[14] A. Kunert, M. Hagemann and N. Erdmann, “Construction of Promoter Probe Vectors for Synechocystis sp. PCC 6803 Using the Light-Emitting Reporter Systems gfp and LuxAB,” Journal of Microbiological Methods, Vol. 41, No. 3, 2000, pp. 185-194. http://dx.doi.org/10.1016/S0167-7012(00)00162-7
[15] O. A. Koksharova and C. P. Wolk, “Genetic Tools for Cyano-bacteria,” Applied Microbiology and Biotechnology, Vol. 58, No. 2, 2002, pp. 123-137. http://dx.doi.org/10.1007/s00253-001-0864-9
[16] A. K. Chaurasia, A. Parasnis and S. K. Apte, “An Integrative Expression Vector for Strain Improvement and Envi-ronmental Applications of the Nitrogen Fixing Cya- no-bacterium, Anabaena sp. strain PCC7120,” Journal of Microbiological Methods, Vol. 73, 2008, pp. 133-141. http://dx.doi.org/10.1016/j.mimet.2008.01.013
[17] N. F. Tsinoremas, A. Kawakami and D. A. Christopher, “High-Fluence Blue Light Stimulates Transcription from a Higher Plant Chloroplast psbA Promoter Expressed in a Cyanobacterium, Synechococcus (sp. Strain CC7942),” Plant and Cell Physiology, Vol. 40, No. 4, 1999, pp. 448- 452. http://dx.doi.org/10.1093/oxfordjournals.pcp.a029562
[18] Y. Yabuta, M. Tamoi, K. Yamamoto, K. Tomizawa, A. Yokota and S. Shigeoka, “Molecular Design of Photosynthesis-Elevated Chloroplasts for Mass Accumulation of a Foreign Protein,” Plant and Cell Physiology, Vol. 49, No. 3, 2008, pp. 375-385. http://dx.doi.org/10.1093/pcp/pcn014

  
comments powered by Disqus

Copyright © 2018 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.