expression and purification of functionally active recombinant human alpha 1-antitrypsin in methylotrophic yeast pichia pastoris
نویسندگان
چکیده
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.
منابع مشابه
Expression and Purification of Functionally Active Recombinant Human Alpha 1-Antitrypsin in Methylotrophic Yeast Pichia pastoris
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 i...
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عنوان ژورنال:
avicenna journal of medical biotechnologyجلد ۳، شماره ۳، صفحات ۱۲۷-۱۳۴
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