КЛОНИРОВАНИЕ И ЭКСПРЕССИЯ кДНК ЭНДО-Β-1,4-ГЛЮКАНАЗЫ ГРИБА ASPERGILLUS NIGER В E. coli И ХАРАКТЕРИСТИКА РЕКОМБИНАНТНОГО БЕЛКА
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Keywords:
cellulase, hydrolysis, recombinant protein,Abstract
EG enzymes are key components in fungal cellulase systems, and their functional activity is critical for hydrolysis of crystalline cellulose. The EG cDNA from A.niger was cloned into E. coli. The method of cloning consisted gene amplification with specific primers. The product of PCR was cloned into E. coli vector plasmid under control of T7 promoter. We showed expression of eng1 gene in E. coli recombinant strain and examined some properties of the recombinant protein. It showed maximal activity at 500C and pH 6.0References
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3.Miyamoto K. Renewable biological systems for alternative sustainable energy production // Food &Agriculture Org., -1997. 108p.
4.Taipakova S.M., Smailov B.B., Stanbekova G., Bissenbaev A. K. Cloning and expression of Lentinula edodes cellobiohydrolase gene in E. coli and characterization of recombinant enzyme // Journal of Cell and
Molecular biology.-Turkey, 2011. -Vol. 9. -№1. -P.53-63.
5.Taipakova S.M., Stanbekova G., Ischenko A., Saparbaev M., Bissenbaev A.K Cloning and expression of Lentinula edodes cellobiohydrolase CEL6B gene in E. coli // International Journal of chemistry and biology. -2011. -№1.-С.19-26.
6. Hon J., Tamaki H., Akiba S., Yamamoto K., Kumagai H. Cloning of Gene Encoding a Highly Stable Endo-β-1,4-Glucanase from Aspergillus niger and Its Expression in Yeast // J. BIOSCI. BIOENG.- 2001.- Vol. 92.-P. 434-441.
7. Nelson NJ. A photometric adaptation of the Somogyi method for the determination of glucose // J Biol Chem. -1944. -Vol.153. –P. 375-380.
8. Somogyi M. Notes on sugar determination // J Biol Chem. -1952. –Vol.195. –P. 19-23.
9. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein using the principle of protein–dye binding // Anal Biochem. -1976. –Vol.72. –P. 248-252.
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Тайпакова, С. М., Жанаева, А. Б., & Бисенбаев, А. К. (2017). КЛОНИРОВАНИЕ И ЭКСПРЕССИЯ кДНК ЭНДО-Β-1,4-ГЛЮКАНАЗЫ ГРИБА ASPERGILLUS NIGER В E. coli И ХАРАКТЕРИСТИКА РЕКОМБИНАНТНОГО БЕЛКА. Experimental Biology, 56(4), 239–244. Retrieved from https://bb.kaznu.kz/index.php/biology/article/view/477
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BIOTECHNOLOGY
