产品货号 | 载体名称 | 出品公司 | 载体用途 | 产品价格 | 出货周期 |
---|---|---|---|---|---|
HG-VJI0571 | pYES2/CT/α-Factor | Invitrogen |
酵母表达载体
|
询价 | 现货 |
质粒类型:酿酒酵母表达载体
诱导方法:半乳糖
启动子:GAL1
5' 测序引物及序列:GAL1-F:AATATACCTCTATACTTTAACGTC
3' 测序引物及序列:CYC1-R:GCGTGAATGTAAGCGTGAC
载体标签:C-His, C-V5
载体抗性:Ampicillin (氨苄青霉素)
筛选标记:URA3
备注:利用半乳糖诱导蛋白在酿酒酵母中表达。
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)。