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

Ashirоv Оybek, Sasmakоv Sobirdjan, Abdurakhmanоv Jaloliddin, HASANОV Shuhrat

Kimyo va kimyo texnologiyasi · 2020-yil

Annotatsiya

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.

Maqola ma’lumotlari
MualliflarAshirоv Оybek, Sasmakоv Sobirdjan, Abdurakhmanоv Jaloliddin, HASANОV Shuhrat
JurnalKimyo va kimyo texnologiyasi
Nashr sanasi2020-01-12
Jild2019
Son4
Tilen
DOI10.70189/1992-9498.1136

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