拟南芥细胞周期调控因子KRP2的原核表达及蛋白纯化
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摘要
表皮毛是由高等植物地上部组织表皮细胞发育而来的一种特殊结构,细胞周期调控因子在植物表皮毛细胞形态的建成过程中发挥着重要作用。利用原核表达系统,构建pET-28a-KRP2-FLAG融合表达载体,在大肠杆菌中对拟南芥细胞周期依赖激酶抑制因子KRP2进行了体外表达,并利用蛋白标签FLAG和His对目的蛋白进行纯化。结果表明,在大肠杆菌系统中成功对KRP2-FLAG进行了体外表达并得到了纯化的蛋白,蛋白量为0.87 mg。这为进一步研究KRP2在表皮毛细胞发育过程中的作用奠定了基础。
引文
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[10]张钰,李亚会,屠唯一,等.拟南芥AtERF1蛋白的原核可溶性表达及多克隆抗体的制备[J].上海师范大学学报(自然科学版),2013,42(4):378-383.
[11]余璐璐,龚绒雪,吕林涛,等.拟南芥氰丙氨酸合酶CYS-C1基因扩增及多克隆抗体制备[J].江苏农业学报,2017,33(6):1235-1241.
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[13]单丽伟,唐如春,刘三阳.小麦种子过氧化物酶WP1基因的原核表达、纯化及多克隆抗体制备[J].生物工程学报,2011,27(1):26-30.
[14]邹向阳,李连宏.细胞周期调控与肿瘤[J].国际遗传学杂志,2006,29(1):70-73.
[2]Menges M,Hennig L,Gruissem W,et al.Cell cycle-regulated gene expression in Arabidopsis[J].Journal of Biological Chemistry,2002,277(44):41987-42002.
[3]Morgan D O.Cyclin-dependent kinases:engines,clocks and microprocessors[J].Annu Rev Cell Dev Biol,1997,13:261-291.
[4]de Veylder L,Beeckman T,Beemster G T,et al.Functional analysis of cyclin-dependent kinase inhibitors of Arabidopsis[J].The Plant Cell,2001,13(7):1653-1668.
[5]Zhou Y M,Wang H,Gilmer S,et al.Effects of co-expressing the plant CDK inhibitor ICK1 and D-type cyclin genes on plant growth,cell size and ploidy in Arabidopsis thaliana[J].Planta,2003,216(4):604-613.
[6]de Clercq A,Inze D.Cyclin-dependent kinase inhibitors in yeast,animals and plants:a functional comparison[J].Crit Rev Biochem Mol Biol,2006,41:293-313.
[7]Verkest A,Manes C L,Vercruysse S,et al.The cyclin-dependent kinase inhibitors KRP2 controls the onset of the endoreduplication cycle during Arabidopsis leaf development through inhibition of mototic CDKA;1 kinase complexes[J].Plant Cell,2005,17(6):1723-1736.
[8]Soichi I,Masaaki U.Cell-cycle control and plant development[J].International Review of Cell and Molecular Biology,2011,291:227-261.
[9]李芳,罗军,许会芬,等.西农萨能羊MAT基因的原核表达及多克隆抗体制备[J].西北农林科技大学学报(自然科学版),2014,42(12):1-6,12.
[10]张钰,李亚会,屠唯一,等.拟南芥AtERF1蛋白的原核可溶性表达及多克隆抗体的制备[J].上海师范大学学报(自然科学版),2013,42(4):378-383.
[11]余璐璐,龚绒雪,吕林涛,等.拟南芥氰丙氨酸合酶CYS-C1基因扩增及多克隆抗体制备[J].江苏农业学报,2017,33(6):1235-1241.
[12]许君,乔帅,常岚,等.苦瓜核糖体失活蛋白MAP30的原核表达与抗体制备[J].河南农业大学学报,2009,43(5):552-556.
[13]单丽伟,唐如春,刘三阳.小麦种子过氧化物酶WP1基因的原核表达、纯化及多克隆抗体制备[J].生物工程学报,2011,27(1):26-30.
[14]邹向阳,李连宏.细胞周期调控与肿瘤[J].国际遗传学杂志,2006,29(1):70-73.