Cloning of cDNA glucoamylase Aspergillus awamori into yeast integrative expression vector

  • Alena V. Kulik Belarusian State University, 4 Niezaliežnasci Avenue, Minsk 220030, Belarus https://orcid.org/0000-0003-4651-7497
  • Olga B. Rus Belarusian State University, 4 Niezaliežnasci Avenue, Minsk 220030, Belarus
  • Anatoliy N. Evtushenkov Belarusian State University, 4 Niezaliežnasci Avenue, Minsk 220030, Belarus
DOI: https://doi.org/10.33581/2521-1722-2019-3-59-66

Abstract

We constructed pKLAC2-based integrative expression plasmid pKGLA-1 with glaA gene from Aspergillus awamori 466. The PCR amplification of the target gene glaA and restriction analysis proved pKGLA-1 construction. Linearised plasmid was used for the integrative transformation of chemically competent Kluyveromyces lactis GG799 cells. Colonies of cells transformed with pKGLA-1 plasmid were selected by growth on agar plates containing 5 mmol/L acetamide. Expression of the heterologous gene in K. lactis cells was visually assessed using medium containing 2 % starch. K. lactis cells containing integrated pKGLA-1 DNA secreted recombinant protein glucoamylase with a native N-terminus.

Author Biographies

Alena V. Kulik, Belarusian State University, 4 Niezaliežnasci Avenue, Minsk 220030, Belarus

PhD (biology); head of the laboratory of transgenic plants at the department of molecular biology, faculty of biology

Olga B. Rus, Belarusian State University, 4 Niezaliežnasci Avenue, Minsk 220030, Belarus

PhD (chemistry), docent; associate professor at the department of molecular biology, faculty of biology

Anatoliy N. Evtushenkov, Belarusian State University, 4 Niezaliežnasci Avenue, Minsk 220030, Belarus

doctor of science (biology), full professor; head of the department of molecular biology, faculty of biology

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Published
2019-10-30
Keywords: cloning, glucoamylase, integrative expression vector, Kluyveromyces lactis
How to Cite
Kulik, A. V., Rus, O. B., & Evtushenkov, A. N. (2019). Cloning of cDNA glucoamylase Aspergillus awamori into yeast integrative expression vector. Experimental Biology and Biotechnology, 3, 59-66. https://doi.org/10.33581/2521-1722-2019-3-59-66
Issue
No 3 (2019): Journal of the Belarusian State University. Biology
Section
Genetics and Molecular Biology

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