摘要
对酵母中FGFR3胞外区蛋白表达进行研究.从pCDNA3.1-myc-His-FGFR3N中酶切得到带有myc/His标签的FGFR3胞外区DNA片段,将其克隆到pYES2中;采用醋酸锂转化法,将重组质粒转入酵母INVSc1;转化子经半乳糖诱导后,提取总蛋白进行Western-blot分析.测序结果表明:pYES2-myc-His-FGFR3N载体构建成功,Western-blot分析证实重组转化子在酵母中表达了与预期分子量大小吻合的FGFR3胞外区蛋白.
Objective ：to study the expression of FGFR3 extracellular domain in yeast. Methods： first getting the myc/His FGFR3 extracellular DNA fragment with restriction enzymes from vector pCDNA3. 1-myc-his-FGFR3N; then, linking the fragment to yeast expressing vector pYES2 and transforming the recombination-vector to yeast INVScl ; and finally, conducting Western-boh analysis after the recombinant protein was induced with galactose. Results： pYES2-FGFR3N DNA sequence analysis shows that the base sequence is correct, Western-blot analysis verifies that the recombinant protein expresses FGFR3 extracellular domain in yeast cell corresponding to the corrected molecular quantity.
出处
《重庆工学院学报（自然科学版）》
2009年第5期45-49,共5页
Journal of Chongqing Institute of Technology
基金
国家重点基础研究发展规划资助项目(973项目)(2005CB522604)
重庆市教委科技项目(KJ070613)
关键词
成纤维细胞生长因子受体3
胞外区
重组
表达
酵母
fibroblast growth factor 3
extracellular domain
recombinant
expression
yeast
