Author/Authors :
Arjmand، Sareh نويسنده National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran , , Bidram، Elham نويسنده Department of Clinical Biochemistry, Tarbiat Modares University , , Sahebghadam Lotfi، Abbas نويسنده , , Shamsara، Mehdi نويسنده , , Mowla، Seyed Javad نويسنده ,
Abstract :
Human alpha 1-antitrypsin (AAT) cDNA was obtained from HepG2 cell lines. After PCR and construction of expression vector pPICZα-AAT, human AAT was expressed in the yeast Pichia pastoris (P.pastoris) in a secretary manner and under the control of inducible alcohol oxidase 1 (AOX1) promoter. The amount of AAT protein in medium was measured as 60 mg/l 72 hr after induction with methanol. Results indicated the presence of protease inhibitory function of the protein against elastase. Purification was done using His-tag affinity chromatography. Due to the different patterns of glycosylation in yeast and human, the recombinant AAT showed different SDS-PAGE patterns compared to that of serum-derived AAT while pI shifted from 4.9 in native AAT compared to 5.2 in recombinant AAT constructed in this study.
