Circular polymerase extension cloning of cDNA glucoamylase Aspergillus awamori into integrative vector for filamentous fungi
https://doi.org/10.29235/1029-8940-2020-65-3-319-327
Abstract
This article describes successful insertion the cDNA glucoamylase from Aspergillus awamori into the integrative vector for filamentous fungi pH4Hyg using the CPEC (circular polymerase extension cloning) strategy. The prior step was the PCR amplification of the cDNA glaA gene cloned into a vector based on the plasmid pBluescript II SK(-) and the amplification of linear pH4Hyg vector with promotor and terminator of the glaA gene. The PCR products were purified and used for CPEC reaction. The CPEC product was formed after 20 cycles of the reaction. It is proposed to use the final plasmid pH4HygPTgla in engineering of filamentous fungi strains producing stable forms of glucoamylase with biotechnological interest.
About the Authors
A. V. Kulik
Belarusian State University
Belarus
Belarus
Alena V. Kulik - Ph. D. (Biol.), Leading Researcher, Belarusian State University.
4, Nezavisimosti Ave., 220030, Minsk.
Yu. V. Selezneva
Belarusian State University
Belarus
Belarus
Yuliya V. Selezneva - Ph. D. (Biol.), Head of the Laboratory, Belarusian State University.
4, Nezavisimosti Ave., 220030, Minsk.
A. V. Kachan
Belarusian State University
Belarus
Belarus
Alexandr V. Kachan - Ph. D. (Biol.), Assistant Professor, Belarusian State University.
4, Nezavisimosti Ave., 220030, Minsk.
O. B. Rus
Belarusian State University
Belarus
Belarus
Olga B. Rus - Ph. D. (Chem.), Assistant Professor, Belarusian State University.
4, Nezavisimosti Ave., 220030, Minsk.
A. N. Evtushenkov
Belarusian State University
Belarus
Belarus
Anatoliy N. Evtushenkov - D. Sc. (Biol.), Professor, Head of the Department, Belarusian State University.
4, Nezavisimosti Ave., 220030, Minsk.
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