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Cloning and screening of the putative hexokinase genes from Rhizopus oryzae and their heterologous expression in Saccharomyces cerevisiae

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Abstract

Background

A filamentous fungus, Rhizopus oryzae (R. oryzae) is one of the ideal candidates for ethanol and lactic acid production due to its ability to grow on renewable carbon sources.

Methods and results

In this study, the nucleotide sequence of hexokinases and glucokinase from S. cerevisiae was found on the NCBI site (http://www.ncbi.nlm.nih.gov/blast/Blast.cgi) were used. With these nucleotide sequences, a blast search was done on the R. oryzae genome database (http://www.broad.mit.edu/annotation/genome/rhizopus_oryzae/Home.html) and ten probable genes were obtained. cDNA was synthesized from the total RNA and PCR products of the seven of these putative genes were determined using the primers designed for them.

Conclusion

The results of the sequences and the complementation studies revealed that three of these seven putative genes were expressed in R. oryzae and the growth was observed on selective media.

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Data availability

This article and its supplementary information files include all the data generated during this study.

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Acknowledgements

This study is supported by TÜBİTAK-BİDEB Ph.D. scholarship (2211) and TÜBİTAK Short Term R&D Funding Program Project Program (1002) (Project No:112T662) and derived from the doctor of philosophy thesis written by Eda ALAGÖZ (Connect to http://etd.lib.metu.edu.tr/upload/12616593/index.pdf ).

Funding

This study is supported by TÜBİTAK-BİDEB Ph.D. scholarship (2211) and TÜBİTAK Short Term R&D Funding Program Project Program (1002) (Project No:112T662).

Author information

Authors and Affiliations

  1. Department of Biotechnology, Middle East Technical University, Ankara, Turkey

    Eda Alagöz, Şeyda Açar, Meral Yücel & Haluk Hamamcı

  2. Department of Biological Sciences, Middle East Technical University, Ankara, Turkey

    Meral Yücel

  3. Department of Food Engineering, Middle East Technical University, Ankara, Turkey

    Haluk Hamamcı

  4. H2 Biyotek, Mustafa Kemal Mahallesi Dumlupınar Bul.No 280 D Blok No:4, Ankara, Turkey

    Haluk Hamamcı

Authors
  1. Eda Alagöz
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  2. Şeyda Açar
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  3. Meral Yücel
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  4. Haluk Hamamcı
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Contributions

All authors contributed to the study's conception and design. ŞA and EA performed material preparation, data collection, and analysis. EA wrote the first draft of the manuscript and all the other authors commented on the previous versions and approved the final manuscript.

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Correspondence to Eda Alagöz.

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Alagöz, E., Açar, Ş., Yücel, M. et al. Cloning and screening of the putative hexokinase genes from Rhizopus oryzae and their heterologous expression in Saccharomyces cerevisiae. Mol Biol Rep 49, 8607–8615 (2022). https://doi.org/10.1007/s11033-022-07696-4

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