CAJ | 학술논문

采用DNA重组技术，构建pGEX-4T-3-CIPK7和pET28-CBL1蛋白表达载体，转化大肠杆菌并诱导表达目的蛋白，利用亲和层析纯化法纯化GST标签、His标签融合蛋白，通过GST pull-down试验验证CIPK7与CBL1之间的相互作用。成功构建了CBL1和CIPK7的重组质粒，经诱导表达及纯化获得了可溶性GST-CIPK7和CBL1-His融合蛋白， GST pull-down试验证实了CBL1能够与CIPK7结合。 CIPK7蛋白与CBL1蛋白之间存在直接的相互作用，为进一步研究蛋白激酶CIPK7的功能奠定了基础。
채용DNA중조기술，구건pGEX-4T-3-CIPK7화pET28-CBL1단백표체재체，전화대장간균병유도표체목적단백，이용친화층석순화법순화GST표첨、His표첨융합단백，통과GST pull-down시험험증CIPK7여CBL1지간적상호작용。성공구건료CBL1화CIPK7적중조질립，경유도표체급순화획득료가용성GST-CIPK7화CBL1-His융합단백， GST pull-down시험증실료CBL1능구여CIPK7결합。 CIPK7단백여CBL1단백지간존재직접적상호작용，위진일보연구단백격매CIPK7적공능전정료기출。
With the DNA recombinant technique, the plasmids pGEX-4T-3-CIPK7 and pET28-CBL1 were constructed. Then the plasmids were transformed into E. coli and induced to express the target proteins. GST and His tagged fusion proteins were purified by affinity chromatography. The binding of CIPK7 and CBL1 was improved via GST pull-down assay. The expression vectors of CIPK7 and CBL1 were successfully constructed, and the soluble GST-CIPK7 fusion protein and CBL1-His tagged protein were obtained via expression and purification. The binding of the two proteins was confirmed by GST pull-down assay. This work proved that there is a direct interaction between CIPK7 and CBL1, and provides some promising clue for the further exploration into the function of CIPK7 kinase.