CAJ | 학술논문

拟南芥的血红蛋白1(AtGLB1)属于非共生的血红蛋白.在低氧胁迫中对植物细胞中过氧化氢(H_2O_2)内稳态的维持起了很重要的作用.为了检测AtGLB1与H_2O_2能否直接相互作用,我们扩增了拟南芥的AtGLB1基因, 并将其克隆到原核表达质粒pET32a中,测序鉴定正确后转化大肠杆菌BL21.IPTG诱导目的蛋白表达后,镍离子亲和层析柱(Ni~(2+) -NTA) 纯化了靶蛋白.体外表达的氧合的AtGLB1能与H_2O_2直接相互作用.因此,与H_2O_2反应可能是AtGLB1清除低氧胁迫下产生的H_2O_2的一种方式.
의남개적혈홍단백1(AtGLB1)속우비공생적혈홍단백.재저양협박중대식물세포중과양화경(H_2O_2)내은태적유지기료흔중요적작용.위료검측AtGLB1여H_2O_2능부직접상호작용,아문확증료의남개적AtGLB1기인, 병장기극륭도원핵표체질립pET32a중,측서감정정학후전화대장간균BL21.IPTG유도목적단백표체후,얼리자친화층석주(Ni~(2+) -NTA) 순화료파단백.체외표체적양합적AtGLB1능여H_2O_2직접상호작용.인차,여H_2O_2반응가능시AtGLB1청제저양협박하산생적H_2O_2적일충방식.
Arabidopsis hemoglobin 1 (AtGLB1) is a kind of non-symbiotic hemoglobin. It plays an important role in sustaining H_2O_2 homeostasis when plant is exposed to hypoxic stress. To detect interaction of AtGLB1 and H_2O_2, the gene encoding AtGLB1 was amplified by PCR, and inserted into prokaryotic expression plasmid pET32a. Then the recombinant expression plasmid pET32a/ AtGLB1 was identified by sequencing and transferred into E.coli BL21. The recombinant protein was expressed under the induction of IPTG, and purified by Ni~(2+) -NTA affinity chromatography column. The recombinant protein could react with H_2O_2 directly. Therefore, reaction with H_2O_2 might be one of ways that AtGLB1 scavenges H_2O_2 produced during hypoxia stress.