| 摘要: |
| 应用基因重组技术,将从华东葡萄白河-35-1中分离出的长1 179 bp芪合酶（stilbene synthase，STS）基因的cDNA片段克隆入大肠杆菌-酵母菌穿梭型诱导表达载体pYES6/CT上，构建重组酵母真核表达质粒pYES6/CT-STS，转化酿酒酵母菌株INVSc1，用Blasticidin（bsd）筛选出阳性克隆转化子，经半乳糖诱导表达后进行5个时间段菌体全蛋白SDS-PAGE电泳和Western-blot分析。结果发现，诱导4 h后开始有目标带（48 ku）出现,诱导16 h开始大量且稳定表达。证明目的基因片段可以在酿酒酵母中表达，为后续基因功能的验证奠定了基础。 |
| 关键词: 芪合酶 酵母表达载体 载体构建 诱导表达 |
| DOI： |
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| 基金项目:国家科技部转基因专项(JY03-A-19-02);西北农林科技大学科研专项（JY-19-02） |
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| Construction of yeast expression vector and inducible expression of cDNA fragment of stilbene synthase in yeast |
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| Abstract: |
| With the technology of gene recombination,we cloned gene of STS full-length cDNA from vitis pesudoreticulata Baihe35-1 into pYES6/CT,a shuttle type plasmid vector of yeast,and transformed INVSc1.The recombinant strains were selected with Blasticidin.The recombinant protein was induced with galactose,and the products of 5 culturing periods were identified by SDS-PAGE and Western blot analysis.The target band was found after induced 4 h,and rich stable expressions of the target protein appeared after 16 h.The result showed that the fragment of target gene from grape could be expressed in yeast,which may lay a foundation for identifying the function of target gene. |
| Key words: stilbene synthase yeast expression vector vector construction expression |
