小麦TaZFP33基因克隆、生物信息学分析、亚细胞定位与表达分析

Cloning,bioinformatics analysis,subcellular localization and expression analysis of TaZFP33 gene from Triticum aestivum

C_(2)H_(2)锌指蛋白作为最大的转录因子家族之一,在调节植物生长发育及逆境胁迫应答反应中起着重要作用。以春小麦品种Fielder(Triticum aestivum L.cv.Fielder)为材料克隆到锌指蛋白TaZFP33基因,其编码区全长510 bp,编码169个氨基酸。生物信息学分析得知:TaZFP33蛋白分子质量为17.5 ku,等电点为6.43,属于Q型锌指蛋白,且为不稳定疏水性蛋白。通过系统进化与启动子分析表明该基因可能受ABA诱导的相关胁迫响应。转化烟草进行瞬时表达分析,结果表明TaZFP33定位在细胞核上。实时荧光定量分析组织特异性表达表明,TaZFP33基因在小麦胚、根、茎、叶中均有表达,其中在胚中表达量最高。经不同浓度的ABA、PEG6000和NaCl处理幼苗后,发现TaZFP33基因对上述胁迫均有不同程度的响应。该研究为后续深入探索TaZFP33基因的生物学功能研究奠定了基础,也为小麦的抗逆性研究提供候选基因。

C_(2)H_(2) zinc finger protein,as one of the largest transcription factor family in plants,plays an important role in plant growth and stress.In this study,the zinc finger protein TaZFP33 gene in the spring wheat variety Fielder(Triticum aestivum L.cv.Fielder)was cloned.The TaZFP33 gene coding region was 510 bp in length and encoded 169 amino acids.Bioinformatics analysis showed that the molecular weight of TaZFP33 protein was 17.5 ku,and the isoelectric point was 6.43,which belonged to the Q-type zinc finger protein and was an unstable hydrophobic protein.Evolutionary analysis and promoter analysis indicated that the gene may respond to ABA-induced stress regulation.Transient expression analysis of transformed tobacco showed that TaZFP33 was localized on the nucleus.Real-time fluorescence quantitative analysis of tissue-specific expression showed that TaZFP33 gene was expressed in stems,leaves,roots and embryos,and the highest expression level was in embryos.After seedlings were treated with different concentrations of ABA,PEG6000 and NaCl,the results showed that TaZFP33 gene responded to above-mentioned stresses.This research laid a foundation for further exploration on the biological function of the TaZFP33 gene,and also provided candidate genes for the study of stress resistance in Triticum aestivum.