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Optimal Conditions Of Fermentation Of The Pichia Pastoris Yeast Expressing The Recombinant Pres2-S Protein (M-Hbsag) Of Hepatitis B Virus (Hbv), Sasmakоv Sobirdjan, Ashirоv Оybek, Abdurakhmanоv Jaloliddin, Xasanоv Shuhrat
Optimal Conditions Of Fermentation Of The Pichia Pastoris Yeast Expressing The Recombinant Pres2-S Protein (M-Hbsag) Of Hepatitis B Virus (Hbv), Sasmakоv Sobirdjan, Ashirоv Оybek, Abdurakhmanоv Jaloliddin, Xasanоv Shuhrat
CHEMISTRY AND CHEMICAL ENGINEERING
The aim of this study is determination of optimal fermentation conditions of the Pichia pastoris yeast expressing the recombinant Pres2-S protein of hepatitis B virus (HBV). For this purpose we investigated the main factors influencing for the cultivation of recombinant strain of the yeast Pichia pastoris pPIC3,5-S-HBsAg - the composition of the nutrient medium, pH and dissolved oxygen (DO). As a result, by enzyme-linked immunosorbent assay (ELISA) the optimal concentration of methanol ≈1.0 %, which is the initiator and the only source of carbon for protein expression in the Mut+ phenotype Pichia pastoris yeast cells was determined. In the selected …
Study Of Recombinant Protein Fractions From The Bombyx Mori And Pichia Pastoris Using "Native Page" Method, Abdurhakhmanov Jaloliddin, Sasmakov Sobirdjan, Khasanov Shuhrat, Ashirov Оybek
Study Of Recombinant Protein Fractions From The Bombyx Mori And Pichia Pastoris Using "Native Page" Method, Abdurhakhmanov Jaloliddin, Sasmakov Sobirdjan, Khasanov Shuhrat, Ashirov Оybek
CHEMISTRY AND CHEMICAL ENGINEERING
We have studied the recombinant protein fractions of ̴ 24-34 kDa obtained from Bombyx mori - silkworm larvae and yeast Pichia pastoris. Fractions, containing the recombinant proteins by gel-electrophoresis using the “Native PAGE” method, which allows for differentiation of proteins according to their natural (surface) charge and conformational properties were studied. The results showed that the recombinant protein fraction from Bombyx mori (̴ 34 kDa) consisted of four components and fraction from Pichia pastoris (̴ 24 kDa) consisted one component. The most optimal conditions for the “Native PAGE”- the main gel: Tris HCl (pH-8.8) 244 mM, Bis-Acrylamide 10%, TEMED 0.08%, …
Study Of Recombinant Protein Fractions From The Bombyx Mori And Pichia Pastoris Using "Native Page" Method, Abdurhakhmanov Aloliddin, Sasmakov Sobirdjan, Khasanov Shuhrat, Ashirov Оybek
Study Of Recombinant Protein Fractions From The Bombyx Mori And Pichia Pastoris Using "Native Page" Method, Abdurhakhmanov Aloliddin, Sasmakov Sobirdjan, Khasanov Shuhrat, Ashirov Оybek
CHEMISTRY AND CHEMICAL ENGINEERING
We have studied the recombinant protein fractions of ̴ 24-34 kDa obtained from Bombyx mori - silkworm larvae and yeast Pichia pastoris. Fractions, containing the recombinant proteins by gel-electrophoresis using the “Native PAGE” method, which allows for differentiation of proteins according to their natural (surface) charge and conformational properties were studied. The results showed that the recombinant protein fraction from Bombyx mori (̴ 34 kDa) consisted of four components and fraction from Pichia pastoris (̴ 24 kDa) consisted one component. The most optimal conditions for the “Native PAGE”- the main gel: Tris HCl (pH-8.8) 244 mM, Bis-Acrylamide 10%, TEMED 0.08%, …
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
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
CHEMISTRY AND CHEMICAL ENGINEERING
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 …