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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Affiliation: | 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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