Cloning of cDNA glucoamylase Aspergillus awamori into yeast integrative expression vector

Author:

Kulik Alena V.1ORCID,Rus Olga B.1,Evtushenkov Anatoliy N.1

Affiliation:

1. Belarusian State University

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.

Publisher

Belarusian State University

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