芒果MiZFP1和MiZFP2基因克隆与表达模式分析

Cloning and Expression Analysis of MiZFP1 and MiZFP2 Genes in Mango

转录因子C2H2型锌指蛋白基因家族(zinc finger proteins,ZFPs)在植物生长发育、逆境胁迫等过程中发挥着重要作用。本研究采用改良RACE技术和RT-PCR技术克隆获得了两个芒果C2H2型锌指蛋白基因全长序列,命名为MiZFP1和MiZFP2。序列分析显示,MiZFP1开放阅读框长度为723 bp,编码241个氨基酸,分子量和等电点分别为25.99 kD和8.70。MiZFP2开放阅读框长度为717 bp,编码239个氨基酸,分子量和等电点分别为25.76 k D和8.31。MiZFP1和MiZFP2的核苷酸和氨基酸序列相似性分别为85%和70%。MiZFP1和MiZFP2氨基酸区域均含有两个高度保守的C2H2型结构域和一个EAR结构域。基因表达模式分析显示,低温胁迫、盐胁迫和干旱胁迫均诱导MiZFP1和MiZFP2基因在叶片和茎中表达,表明MiZFP1和MiZFP2基因可能在芒果逆境胁迫应答中起重要作用。本研究成功将MiZFP1和MiZFP2基因完整编码区序列导入植物表达载体pBI121,可为下一步深入开展该基因功能研究提供理论依据。

The transcription factors C2H2-type zinc finger proteins(ZFPs)family play essential roles in plant developmental control and various abiotic stresses responses.In present study,two full-length C2H2 zinc finger proteins were cloned by modified RACE technology and RT-PCR technology in Mangifera indica L.(mango),named MiZFP1 and MiZFP2.Sequences analysis showed that the open reading frame of MiZFP1 was 723 bp in length,coding 241 amino acids protein,the molecular weight and isoelectric point(pI)were 25.99 kD and 8.70,respectively.The MiZFP2 open reading frame length was 717 bp,encod 239 amino acids protein of 25.76 kD with a pI of 8.31.The nucleotide and amino acid sequences homology of MiZFP1 and MiZFP2 were 85%and 70%respectively.The amino acid regions of MiZFP1 and Mi ZFP2 both contain two high conserved C2H2-type domains and one EAR domain.Expression analysis showed that low temperature,salt and drought stress induce MiZFP1 and MiZFP2 expression in leaves and stems.The results indicated that MiZFP1 and MiZFP2 may play an important role in mango stress response.The full coding sequences of MiZFP1 and MiZFP2 were successfully inserted into the plant expression vector pBI121.This work provides a theoretical basis for further research on the function of these genes.