拟南芥高迁移率族蛋白B族基因At2G34450在毕赤酵母中的表达及纯化

摘要

[Objective] The aim was to study the expression of Arabidopsis gene A/2G34450 in Pichia pastoris and to obtain recombinant Arabidopsis HMGB protein. [Method] The At2G34450 gene was cloned into yeast expression vector pPIC9K containing AOXl promoter and the sequences of secreting α-signal peptides. Recombinant plasmid was linearized by Sal l and transformed into P. pastoris GSl15 competent cells by electroporation. Positive integrated clones were screened out, and the At2G34450 protein was expressed under the induction of methanol. [Result] The At2G34450 protein was expressed in yeast medium through methanol induction. SDS-PAGE results showed that recombination product was At2G34450 protein. [Conclusion] At2G34450 protein was successfully expressed in the P. pastoris system for the first time, which paves a direct path to further research on the functions of HMGB family members.%观察拟南芥高迁移率族蛋白B族基因At2G34450在毕赤酵母体系中的表达，获得重组蛋白。[方法]将At2G34450基因插入含AOX1启动子和d分泌信号肽序列的酵母表达载体pPIC9K中，用SalI将重组质粒线性化，电击转化毕赤酵母GS115感受态细胞，筛选阳性整合子进行甲醇诱导表达。[结果]拟南芥At2G34450在酵母培养基中实现了表达，表达产物经SDS—AGE鉴定为重组蛋白。[结论]在毕赤酵母真核系统中实现了拟南芥At2G34450蛋白的表达，为进一步研究拟南芥HMGB家族蛋白打下了基础。