拟南芥AtXCD1蛋白的原核表达研究Prokaryotic Expression of Arabidopsis AtXCD1 Fusion Proteins
陈光朗;董万春;韩娇;樊婷婷;杨立波;曹树青;
摘要(Abstract)：
[目的]研究拟南芥AtXCD1蛋白的原核表达。[方法]构建XCD1原核表达载体,根据NCBI基因序列设计引物,并以PCR扩增得到大量目的片段XCD1,将XCD1连接到原核表达载体pET-32a+,并对重组质粒进行测序鉴定,将重组质粒转化至大肠杆菌原核表达菌株BL21(DE3),对蛋白表达条件:诱导时间、诱导温度和IPTG浓度等进行优化。IPTG诱导表达获得目的蛋白,对蛋白表达条件:诱导时间、诱导温度和IPTG浓度等进行优化。[结果]XCD1序列全长为1 242 bp,与PCR产物大小一致。蛋白XCD1-pET-32a+较适表达条件为在30℃0.1 mmol/L的IPTG诱导1.5 h。[结论]该结果为进一步蛋白纯化及酶活测定试验奠定了基础。
关键词(KeyWords)： 拟南芥(Arabidopsis thaliama);AtXCD1;重组载体;原核表达
基金项目(Foundation): 安徽省青年科学基金项目(1308085QC56);; 高等学校博士学科点基金(20120111110009)
作者(Author): 陈光朗;董万春;韩娇;樊婷婷;杨立波;曹树青;
Email:
DOI: 10.13989/j.cnki.0517-6611.2014.14.006
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