细胞壁转化酶(cell wall invertase,CWI)催化质外体蔗糖水解形成源/库的糖浓度梯度,促进蔗糖不断地向库细胞卸出,在作物库强调节中起着十分关键的作用。为揭示小麦(Triticum aestivum)CWI在籽粒灌浆期的重要作用,本研究对已克隆的小麦细胞壁转化酶基因(Ta CWI-B1)进行了生物信息学分析,并利用Ta CWI-B1的编码区构建了原核表达载体p ET28a-Ta CWI-B1,转化大肠杆菌(Escherichia coli)BL21(DE3)菌株后对异丙基硫代半乳糖苷(isopropylβ-D-1-thiogalactopyranoside,IPTG)诱导浓度、培养温度和培养时间进行了摸索;利用十二烷基硫酸钠聚丙烯酰胺凝胶电泳(sodium dodecyl sulfate polyacrylamide gel electrophoresis,SDS-PAGE)分析重组蛋白可溶性,利用镍柱分离纯化蛋白后通过基质辅助激光解吸电离飞行时间质谱(matrix-assisted laser desorption/ionization time of flight mass spectro...
【英文摘要】
Cell wall invertase(CWI) is the key regulator which forms sugar concentration gradient between source and sink by catalyzing sucrose hydrolysis and promotes sucrose into sink. In order to uncover the important roles of CWI in wheat(Triticum aestivum), bioinformatics were utilized to analyse the protein properties and structure map of wheat Ta CWI-B1.The prokaryotic expression vector p ET28a-Ta CWI-B1 was constructed by inserting the coding region of Ta CWI-B1 into p ET28a,and transformed into Escherichia co...
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