pYES2/CT/α-Factor产品信息

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
HZ0571	pYES2/CT/α-Factor		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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