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Molecular Cloning and Prokaryotic Expression of the Gene Encoding HMG-CoA Reductase from Eucalyptus urophylla×E.grandis

尾巨桉3-羟基-3-甲基戊二酰CoA 还原酶基因的克隆及表达分析


The gene encoding 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) was cloned by Rapid Amplification of cDNA Ends (RACE) method from the leaf tissue of Eucalyptus urophylla × E. grandis . The gene was designated EuHMGR1 and its GenBank accession number is GQ915611. The full-length EuHMGR1 cDNA was 1 955 bp, containing a 1 560 bp open reading frame(ORF) which encodes a peptide of 519 amino acids with a calculated molecular mass of 55 ku. The predicted EuHMGR1 showed 74%,72%,72%,70% and 70% identities with Cyclocarya paliurus,Gossypium hirsutum,Hevea brasiliensis,Tilia miqueliana and Nicotiana tabacum by Blast P on-line NCBI. Tissue expression profile analysis by Real-Time quantitative PCR indicated EuHMGR1 expression was the highest in branch, followed by leaf and no expression in root. Through constructing the prokaryotic expression vector pQE-30-EuHMGR1 into Escherichia coli M15 strain, the recombinant EuHMGR1 protein about 55 ku was successfully obtained by IPTG induction.


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