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Constitutive Expression of α-galactosidase Mel1 Gene in Pichia pastoris
ZHANGXue-wen;TANGXiang-shan;ZHANGJin-chen;ZHANGHuai-yun
College of Bio-science and Technology, Hunan Agricultural University, Changsha 410128, China;
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Abstract  Mel1 gene was amplified from yeast(Saccharomyces cerevisiae ) AH109 with PCR, and cloned into integrated vector pGAPZαA, constitutive and secreted expression α-galactosidase plasmid pGAPZα-Mel1 was constructed. The linearized recombinant plasmid pGAPZ α-Mel1 was transformed into Pichia pastoris KM71 by electropotation, and the blue positive colonies were screened out on YPDS plate with X-α-gal and 100 mg/mL zeocin. There was a special 53 kD band by SDS-PAGE from supernatant of yeast culture and a color band was observed by soak the native PAGE gel in X-α-gal to illustrate the α-galactosidase activity; The α-galactosidase enzyme activity was 12 U/mL after fermenting pGAPZα-Mel1 /KM71 for 6 days.
Key wordsα-galactosidase      Mel1 gene      recombinant plasmid pGAPZα-Mel1      constitutive expression     
Received: 06 April 2005     
Corresponding Authors: ZHANG Xue- wen   
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ZHANGXue-wen
TANGXiang-shan
ZHANGJin-chen
ZHANGHuai-yun
Cite this article:   
ZHANGXue-wen,TANGXiang-shan,ZHANGJin-chen, et al. Constitutive Expression of α-galactosidase Mel1 Gene in Pichia pastoris[J]. , 2006, 14(2): 269-272.
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http://journal05.magtech.org.cn/Jwk_ny/EN/      OR     http://journal05.magtech.org.cn/Jwk_ny/EN/Y2006/V14/I2/269
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