尾巨桉HMGR基因的克隆及表达分析

Molecular Cloning and expression analysis of 3 hydroxy 3 methylglutaryl CoA Reductase Gene from Eucalyptus urophylla×E.grandis

关键词:  尾巨桉  HMGR  生物信息学  内含子  组织特异性表达

Key words:  Eucalyptus urophyla×E.grandis  HMGR  bioinformatics  intron  tissue specific expression

摘要:

基于RACE技术克隆得到尾巨桉3 羟基 3 甲基戊二酰CoA还原酶（EuHMGR）基因全长为1 955 bp，包含1 560 bp的开放阅读框（ORF），编码519个氨基酸。生物信息学分析表明EuHMGR包含两个HMG CoA结合基序和两个NADP(H)结合基序；同源建模得到EuHMGR三维构象呈“V”型，包含N 结构域、S 结构域和L 结构域。将EuHMGR组DNA与cDNA序列进行比对表明EuHMGR存在一个319 bp的内含子。通过实时定量PCR进行组织特异性表达分析表明EuHMGR在茎中的表达量最高，叶次之，在根中基本无表达。该研究为后续尾巨桉EuHMGR的功能验证以及遗传转化奠定基础。

Abstract:

The gene encoding 3 hydroxy 3 methylglutaryl CoA reductase (HMGR)was cloned by Rapid Amplification of cDNA Ends (RACE)method，the full length EuHMGR cDNA was 1955 bp，containing a 1 560 bp open reading frame，encoding a peptide of 519 amino acids. Bioinformatics analysis indicated EuHMGR contained two HMG CoA binding motifs and two NADP(H)binding motifs；and it had a 3 D structure with“V”homology based on modeling，containing N domain，S domain and L domain. There was a 319 bp intron in EuHMGR gene genomic DNA sequence compared with the cDNA sequence. Tissue expression profile analysis by Real Time quantitative PCR indicated EuHMGR expression was the highest in stem，followed by leaf 〖JP3〗and no expression in root. The paper would supply some information to the function analysis and genetic transformation of EuHMGR from Eucalyptus urophylla×E.grandis.