摘要
以‘富平尖柿’(DiospyroskakiL.‘FupingJianshi’)果实不同发育时期提取的总RNA为模板,利用RT-PCR结合RACE技术扩增得到5个扩张蛋白基因:成熟期CDK-Exp3(1168bp),转色期CDK-Exp4(1120bp)和CDK-Exp5(1018bp),膨大期CDK-Exp6(1129bp)和CDK-Exp7(1121bp);5个基因的开放阅读框(ORF)均为765bp,编码254个氨基酸;构建了CDK-Exp3原核表达载体pET-CDK-Exp3,并转化于E.coliBL21(DE3),SDS-PAGE检测结果显示表达了一个约43kD的蛋白。
Using the total RNA from different stages of persimmon（Diospyros kaki L.＇Fuping Jianshi＇）fruit as the template,five expansin cDNA：CDK-Exp3（1 168 bp）from mature stage,CDK-Exp4（1 120 bp）and CDK-Exp5（1 018 bp）from colour-changed stage,CDK-Exp6（1 129 bp）and CDK-Exp7（1 121 bp）from fruit expanding stage were isolated by RT-PCR and RACE.The open reading frame of the five genes was 765 bp and encoded a protein of 254 amino acid residues.The CDK-Exp3 was cloned into pET-32a（＋）vector to construct recombination prokaryotic expression vector pET-CDK-Exp3,and which was transformed to E.coli BL21.Recombinant protein about 43 kD was expressed in pET-CDK-Exp3 system and separated by SDS-PAGE electrophoresis.
出处
《园艺学报》
CAS
CSCD
北大核心
2010年第7期1139-1146,共8页
Acta Horticulturae Sinica
基金
国家自然科学基金项目(30771756)
