摘要
以已构建的含有小麦低分子量麦谷蛋白基因XYGluD3-LMWGS1的重组质粒pGEM-XYGluD3-LMWGS1为模板,利用设计的序列特异引物,扩增基因XYGluD3-LMWGS1编码区,构建成巴斯德毕赤酵母表达载体pPIC3.5K-XYGluD3-LMWGS,将其线性化后用电激法导入甲醇型酵母(Pichia.pastoris)菌株GS115中;利用pPIC3.5K特征引物进行PCR鉴定,筛选得到重组转化子,将重组转化子进行蛋白诱导表达,经SDS-PAGE电泳检测,XYGluD3-LMWGS1基因未能在毕赤酵母中成功表达。
The conding region of XYGluD3-LMWGS1 gene was amplified from the recombinant plasmid pGEM-XYGluD3-LMWGS1 using specific primers, and cloned into pPIC3. 5K, the expressive vector of Pichia. pastoris. The results indicated that the XYGluD3-LMWGS1 gene was cloned into pPIC3.5K, and the expressive vector was transformed into Pichia pasteoris （GSl15） by electricity dash method and the recombinants were identified by the PCR. Then the recombinants was induced for target protein expressing. However, SDS-PAGE showed that XYGluD3-LMWGS1 gene was not expressed in Pichia pastoris.
出处
《西北农业学报》
CAS
CSCD
北大核心
2007年第6期67-71,共5页
Acta Agriculturae Boreali-occidentalis Sinica
基金
西北农林科技大学青年学术骨干支持计划课题
科技部转基因植物研究与开发专项课题(JY-03-13-1)
