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尾巨桉HMGR基因的克隆及表达分析
蒋春1, 张华玲2, 彭江2*   
1.四川大学 生命科学学院 生物资源与生态环境教育部重点实验室, 四川 成都 610064;2.2.西南大学 生命科学学院 三峡库区生态环境教育部重点实验室, 重庆 400715
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。
关键词:  尾巨桉  HMGR  生物信息学  内含子  组织特异性表达
DOI:
分类号:Q943.2;Q946.85
Fund project:
Molecular Cloning and expression analysis of 3 hydroxy 3 methylglutaryl CoA Reductase Gene from Eucalyptus urophylla×E.grandis
JIANG Chun1, ZHANG HuaLing2, PENG Jiang2*   
1.Key Laboratory of Bioresources and Ecoenvironment of Ministry of Education, School of Life Sciences, Sichuan University, Chengdu 610064, China;2.Key Laboratory of Ecoenvironments in Three Gorges Reservoir Region, Ministry of Education, School of Life Sciences, Southwest University, Chongqing 400715, China
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.
Key words:  Eucalyptus urophyla×E.grandis  HMGR  bioinformatics  intron  tissue specific expression
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