摘要
WRKY转录因子是植物所特有的一个超级基因家族,能调控植物生长发育过程及逆境反应。根据转录组测序获得的EST序列设计引物,利用RT-PCR及RACE技术克隆了甘蓝型油菜WRKY30基因cDNA全长序列,并运用生物信息学手段对序列进行了分析及预测。结果表明:BnaWRKY30-like基因全长cDNA序列中包含102 bp的5′-UTR序列,188 bp的3′-UTR序列以及942 bp的完整ORF,编码313个氨基酸。该转录因子蛋白相对分子量为35319.85D,等电点为6.22,基本氨基酸中含量最高的丝氨酸(Ser),313个氨基酸中含有41个酸性氨基酸,34个碱性氨基酸,在121~182区域含有一个典型的WRKY结构域。亚细胞定位预测发现该转录因子定位于细胞核的可能性最高。蛋白质的二级结构预测表明共有α-螺旋占26.20%,延伸链占10.54%,β-转角占4.47%,无规则卷曲占58.79%。BnaWRKY30-like基因编码的蛋白具有典型的WRKY转录因子特征,与拟南芥WRKY30基因亲缘关系近,可能在油菜早期生长发育过程及盐胁迫中的发挥调控作用。
WRKY transcription factor is a unique super gene family of plants,which can regulate the growth and development process and stress response of plants.Based on the EST sequence obtained from transcriptome sequence,the full-length sequence of WRKY30 gene in Brassica napus was cloned by RT-PCR and RACE technology,and the sequence was analyzed and predicted by bioinformatics.Sequence analysis showed that the full-length cDNA sequence of BnaWRKY30-like gene contained 102 bp 5’-UTR,188 bp 3’-UTR and 942 bp ORF,encoding 313 amino acids.The protein has a relative molecular weight of 35319.85 D,isoelectric point of 6.22,serine(Ser)with the highest content in basic amino acids,41 acidic amino acids and 34 basic amino acids,and a typical WRKY domain in 121-182 region.Subcellular localization predicted that the transcription factor was most likely to be located in the nucleus.The prediction of protein secondary structure showed that there were 26.20% of α-helix,10.54%of extended strand,4.47% of β-turn and 58.79% of random coil.The encoded protein of BnaWRKY30-like gene is a typical WRKY transcription factor,which is closely related to Arabidopsis WRKY30 gene,and may play a regulatory role in the early growth and development and salt stress.
作者
尹明智
胡燕
YIN Mingzhi;HU Yan(College of Biology and Agricultural Science&Technology,Zunyi Normal University,Zunyi 563006)
出处
《安徽农业大学学报》
CAS
CSCD
2020年第5期832-838,共7页
Journal of Anhui Agricultural University
基金
贵州省科学技术厅、遵义市科学技术局、遵义师范学院联合科技基金资助(黔科合LH字[2016]7016号)
贵州省高层次创新型人才培养计划项目(遵市科合人才[2017]11号)
遵义师范学院2017年学术新苗培养及创新探索项目培育项目(黔科合平台人才[2017年]5727-15号)共同资助。
