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pYES2/CT/α-Factor产品信息

订购信息

货号 载体名称 出品公司 载体用途 价格 到货周期
VJI0571 pYES2/CT/α-Factor Invitrogen 酵母表达载体 1000
900
现货

产品参数

载体用途:酿酒酵母表达载体

诱导方法:半乳糖

原核抗性:Ampicillin(氨苄青霉素)

筛选标记:URA3

启动子:GAL1 promter

标签:C端带V5标签及His标签

5' 测序引物:GAL1-f(5'-AATATACCTCTATACTTTAACGTC-3')

3' 测序引物:CYC1-r(5'-GCGTGAATGTAAGCGTGAC-3')

备注:利用半乳糖诱导蛋白在酿酒酵母中表达。

载体描述

pPIC9 was used as a template and α-factor gene of Saccharomyces cerevisiae was amplified by PCR. The gene was cloned in the intracellular expression vector pYES2/CT for Saccharomyces cerevisiae and the secreting expression vector named pYES2/CT/α-factor(pYCα) was constructed. The gene of mannase(man) from recombinant vector of pKLAC1-man (pKLman) was cut by restriction enzymes and linked with pYCα. This recombinant vector pYCα-man was used to determine the secretory ability and stability of pYCα. The excellent secretory ability of pYCα was proved by two experiments. One showed that INVSc1/pYCα-man clones formed the clear rings around the clones on the medium contained trypan blue, while INVSc1/pYCα clones had no rings.Further analysis of mannase activity of extracellular supernatant and intracellular extracts showed that both extracellular and intracellular mannase activities of INVSc1/pYCα were not detected,while INVScl/pYCα-man had evident extracellular mannase activities and no intracellular mannase activities. The stability of pYCα was also very good proved by continuous cultivation for about 150h.

 

以pPIC9为模板,通过PCR扩增获得酿酒酵母的α-交配因子(α-factor),并克隆至酿酒酵母胞内表达载体pYES2/CT中,构建了一种新型酿酒酵母附加型分泌表达载体pYES2/CT/α-factor(pYCα)。

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