关键词: 尾巨桉, 3-羟基-3-甲基戊二酰CoA还原酶, RACE, 组织特异性表达, 原核表达

Abstract:

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.

Key words: Eucalyptus urophylla×, E.grandis, HMGR, RACE, tissue specific expression, prokaryotic expression