Obtaining of the “green flourecent protein - gfp” in yeast of Pichia pastoris as repoter of heterological protein expression

Authors

ASHIRОV Оybek, Institute of the Chemistry of Plant Substances, Tashkent, UzbekistanFollow
SASMAKОV Sobirdjan, Institute of the Chemistry of Plant Substances, Tashkent, UzbekistanFollow
ABDURAKHMANОV Jaloliddin, Institute of the Chemistry of Plant Substances, Tashkent, UzbekistanFollow
HASANОV Shuhrat, Institute of the Chemistry of Plant Substances, Tashkent, UzbekistanFollow

Abstract

Based on pPIC3.5 and pPIC9 yeast vectors the recombinant plasmid DNA pPIC3.5-GFP (8464 bp) and pPIC9-GFP (8718 bp) containing cDNA (719 bp) of Green fluorescent protein (GFP) were cloned. The obtained plasmids were transformed into yeast cells of the GS115 Pichia pastoris strain. Transformants are selected on histidine-deficient medium. Recombinant clones of pPIC3.5-GFP and pPIC9-GFP were identified by detecting cell fluorescence. GFP expression in yeast is confirmed by green fluorescence. Using electrophoresis in PAAG the molecular weight of the synthesized protein (≈27 kDa) is determined. The resulting reporter constructs and recombinant strains can be used to study the expression level of various recombinant proteins in Pichia pastoris cells.

Recommended Citation

Оybek, ASHIRОV; Sobirdjan, SASMAKОV; Jaloliddin, ABDURAKHMANОV; and Shuhrat, HASANОV (2019) "Obtaining of the “green flourecent protein - gfp” in yeast of Pichia pastoris as repoter of heterological protein expression," CHEMISTRY AND CHEMICAL ENGINEERING: Vol. 2019: No. 4, Article 27.
Available at: https://cce.researchcommons.org/journal/vol2019/iss4/27