| 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 |
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| 引用本文： | 肖庆振,王日文,王洪霞,冀芦沙.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[J].安徽农业科学,2011,39(32):19674-19676,19683. |
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| 作者姓名： | 肖庆振 王日文 王洪霞 冀芦沙 |
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| 作者单位： | 聊城大学生命科学学院,山东聊城,252059;聊城大学生命科学学院,山东聊城,252059;聊城大学生命科学学院,山东聊城,252059;聊城大学生命科学学院,山东聊城,252059 |
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| 基金项目： | 聊城大学博士启动基金项目(31805) |
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| 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。
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| 关 键 词： | 拟南芥 高迁移率族蛋白 毕赤酵母 真核表达 |
Expression and Purification of the Arabidopsis High Mobility Group B Proteins gene At2G34450 in Pichia pastoris |
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| Institution: | XIAO Qing-zhen et al(School of Life Science,Liaocheng University,Liaocheng,Shandong 252059) |
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| Abstract: | 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. |
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| Keywords: | Arabidopsis High mobility group protein Pichia pastoris Eukaryotic expression |
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