| 刘智黄志刚吴尚虹彭琼于峰.拟南芥生长素结合蛋白ABP1 的原核表达及蛋白纯化[J].,2012,12(26):5049-5052 |
| 拟南芥生长素结合蛋白ABP1 的原核表达及蛋白纯化 |
| Prokaryotic Expression and Protein Purification of the Auxin BindingProtein 1 in Arabidopsis |
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| DOI： |
| 中文关键词: 拟南芥 生长素结合蛋白 原核表达 蛋白纯化 |
| 英文关键词: Arabidopsis Auxin Binding Protein Prokaryotic Expression Protein Purification |
| 基金项目:湖南农业大学科学人才科学基金项目（08YJ06） |
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| 中文摘要: |
| 目的：鉴于生长素结合蛋白(Auxin Binding Protein，ABP)能与生长素特异性结合，因而探讨研究其直接用于生长素信号转导
机理和生物传感器的可能性与可行性。方法：通过RT-PCR 获得拟南芥生长素结合蛋白1 (Auxin bing protein 1, ABP1) 的全长
CDS，将其克隆到原核表达载体pGEX4T-1中，成功构建pGEX4T-1-ABP1重组表达载体。经酶切、PCR及DNA测序鉴定后，将阳
性质粒转化表达受体菌BL21(DE3)。加入异丙基-β-D-硫代半乳糖苷(IPTG)进行诱导后，取样进行SDS-PAGE分析。结果：成功表
达出一个分子量约为43 kD的可溶性融合蛋白，并利用GST亲和柱纯化方式得到了ABPl。结论：通过原核表达并经GST柱纯化
后获得ABP1，为生长素生物传感器的研制开辟新的途径。同时为进一步研究ABP1与生长素的信号转导机制和生长素在生物传
感测定技术中的研究和应用奠定基础。 |
| 英文摘要: |
| Objective: Auxin-binding protein can bind an auxins specifically, and can be used to study the mechanism of the signal
transduction of auxin and bio-sensors. Methods：An auxin-binding protein gene (Auxin bing protein 1, ABP1) was obtained by RT-PCR
from arabidopsis, and was cloned into the prokaryotic expression vector pGEX4T-1（pGEX4T-1-ABP1）. Identified by digestion, PCR
and DNA sequencing, the recombinant plasmid was transformed into the foreign bacteria BL21(DE3). The ABP1 protein was induced by
IPTG and detected by SDS-PAGE. Results: The soluble fusion protein was expressed successfully with a molecular weight of 43 kDa.
The protein was purified with GST affinity column. Conclusion：The prokaryotic expression and purity of GST-ABP1 protein show a way
to development new auxin biosensor and also laid a foundation for the further study of auxin signal transduction and biosensor research. |
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