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  

Expression and Purification of the Arabidopsis High Mobility Group B Proteins gene At2G34450 in Pichia pastoris

作　　者：肖庆振[1] 王曰文[1] 王洪霞[1] 冀芦沙[1] 

机构地区：[1]聊城大学生命科学学院,山东聊城252059

出　　处：《安徽农业科学》2011年第32期19674-19676,19683,共4页Journal of Anhui Agricultural Sciences

基　　金：聊城大学博士启动基金项目(31805)

摘　　要：[目的]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α分泌信号肽序列的酵母表达载体pPIC9K中,用SaⅠl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-PAGE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。[Objective]The aim was to study the expression of Arabidopsis gene At2G34450 in Pichia pastoris and to obtain recombinant Arabidopsis HMGB protein.[Method]The gene At2G34450 was cloned into yeast expression vector pPIC9K containing AOXI promoter and the sequences of secreting α-signal peptides.Recombinant plasmid was linearized by SalI and transformed into P.pastoris GS115 competent cells.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 products were At2G34450 protein.[Conclusion]At2G34450 protein successfully achieves expression in the P.pastoris system at first time,which pave a direct path to further research on function of HMGB family members.

关 键 词：拟南芥 高迁移率族蛋白 毕赤酵母 真核表达 

分 类 号：S132[农业科学—农业基础科学]