作物学报 ›› 2015, Vol. 41 ›› Issue (05): 717-724.doi: 10.3724/SP.J.1006.2015.00717
成伟,郑艳茹,葛丹凤,程光远,翟玉山,邓宇晴,彭磊,谭向尧,徐景升*
CHENG Wei,ZHENG Yan-Ru,GE Dan-Feng,CHENG Guang-Yuan,ZHAI Yu-Shan,DENG Yu-Qing,PENG Lei,TAN Xiang-Yao,XU Jing-Sheng*
摘要:
植物在受到低温、高盐、干旱等非生物逆境胁迫后,CBF结合因子(C-repeat/dehydration-responsive element binding factor)会诱导一系列非生物胁迫应答基因的表达,在提高植物抗逆性方面具有重要作用。本研究利用生物信息学和RT-PCR (reverse transcription-polymerase chain reaction)方法从甘蔗中克隆到1个新的CBF类基因,命名为ScCBF1。该基因的开放读码框(open reading frame, ORF)长度为603 bp,编码200个氨基酸,编码蛋白相对分子质量为22.80 kD,理论等电点为10.31。氨基酸序列比对结果表明,ScCBF1与高粱(Sorghum bicolor)和玉米(Zea mays)中该蛋白相似度分别是96%和94%。进化分析表明ScCBF1与高粱的亲缘关系最近。qRT-PCR表达分析结果表明,ScCBF1在甘蔗根、茎、叶中均有表达,但在根中表达量最高。ScCBF1在低温、干旱、脱落酸(ABA)胁迫下诱导表达,而高盐抑制ScCBF1的表达。成功构建了原核表达载体pGEX-6P-1-ScCBF1,通过IPTG诱导,实现了ScCBF1蛋白在大肠杆菌中的表达。
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