小麦泛素融合降解蛋白基因全长cDNA的克隆及分析

Cloning and Analysis of Ubiquitin Fusion Degradation Protein GenecDNA in Wheat

实验利用RT PCR技术,在小麦矮苏3品种中克隆了1个编码泛素融合降解蛋白基因的cDNA,并且含有完整的5′端,将该基因命名为Tufd1,利用RACE技术克隆了该cDNA的3′端。根据这2段cDNA克隆,设计特异引物,利用RT PCR扩增出了Tufd1完整的开放读码框(ORF),其编码区长948bp,编码315个氨基酸的多肽,在NCBI中运行BLAST。分析表明,Tufd1蛋白同拟南芥的UFD1蛋白有74%的同源性,在所编码的多肽链的N 端有UFD1保守结构域,可作为催化蛋白降解的信号。

Ubiquitin fusion degradation protein is involved in a degradation pathway for ubiquitin fused products,firstly described in yeast.A full-length cDNA encoding the ubiquitin fusion degradation protein was cloned through RT-PCR and RACE,and this gene was designated as Tufd1.The entire coding region of Tufd1 was amplified by using special primers,which was 948bp,encoding a polypeptide of 315 amino acids.Its N-terminal has a conserved UFD1 domain functioning as a degradation signal of proteins.This gene was first cloned in wheat genomes.