摘要： 为了研究强抗逆植物沙冬青寒旱诱导表达基因AmZFP1（Zinc finger protein 1）的生理功能，通过转录谱分析，从中鉴定出一个受寒旱诱导的ZFP全长cDNA序列并命名为AmZFP1。利用半定量RT-PCR方法对AmZFP1进行了表达分析，结果表明，在室内正常培养的沙冬青幼苗中该基因有较高表达量，在低温或干燥处理2~48 h其表达量明显上调，尤其以干燥处理上调速度更快且上调幅度略高；在野外自然生长植株的嫩叶、花蕾和幼嫩果荚中AmZFP1均有较明显的表达，而在嫩枝和根中检测不到其转录本；在野外植株的嫩叶中，AmZFP1在严冬季节高表达，而在春、夏、秋温热季节表达水平低或很微弱。利用RT-PCR方法克隆到AmZFP1编码区cDNA（816 bp），推测其编码蛋白含有2个典型C2H2型锌指结构域及一个核定位信号。此外成功将该cDNA片段构建到植物表达载体p3300-353T上，为下一步深入开展该基因功能研究奠定了基础。

关键词: 沙冬青, 锌指蛋白, 表达分析, 基因克隆

Abstract: To investigate the physiological function of a cold-and drought-inducible gene AmZFP1 (Zinc finger protein 1) from Ammopiptanthu smongolicus with strong stress tolerance,the expression analysis of AmZFP1 was performed by using the semi-quantitative RT-PCR method.The results showed that in the seedlings cultivated in normal growth conditions,a relatively high expression level of AmZFP1 was detected.After exposing the seedlings to low temperature or dehydration treatment between 2 to 48 hours,the expression level of AmZFP1 increased markedly,and this up-regulation was more rapid and slightly higher during the dehydration treatment.In the young leaves,flower buds and immature pods of the A. mongolicus plants grown in the wild,the expression levels of AmZFP1 were quite high,whereas in the twigs and roots,the transcripts of AmZFP1 were not detected.In young leaves of the wild grown A. mongolicus plants,the expression level of AmZFP1 was obviously high in the cold winter but was low or quite weak in the warm or hot seasons,including spring,summer and autumn.Moreover,the coding region cDNA of AmZFP1 was cloned by RT-PCR,whose predicted polypeptide contains two typical C2H2 type zinc finger domains and one nuclear localization signal.The cDNA fragment was also successfully inserted into the plant expression vector p3300-35ST.This work laid a foundation for further functional analysis of AmZFP1.

Key words: Ammopiptanthus mongloicus, ZFPs, Expression analysis, Gene cloning

中图分类号: 

• Q78