关键词: 油桐, 生物素羧基载体蛋白(BCCP), 基因克隆, 表达分析

Abstract:

Biotin carboxyl carrier protein (BCCP) plays an important role as a bridge link in the reaction of Acetyl CoA carboxylase. A full-length cDNA sequence of BCCP gene was obtained from kernel cDNA library of Vernicia fordii, named VfBCCP (GenBank accession No.JQ736807). Sequence analysis indicated that VfBCCP was 1 023 bp in full length with a 5'-untranslated region(5'-UTR) of 82 bp, a 3'-UTR of 182 bp, an opening reading frame(ORF) of 759 bp which encoding 252 predicted amino acids. Furthermore, the genome sequence of BCCP was isolated with 2 157 bp in length which including 5 introns (GenBank accession No.JX171700). The amino acid sequence alignment and phylogenetic analysis revealed that VfBCCP had the closest phylogenetic relationship with that of Jatropha curcas belonging to Euphorbiaceae and they shared 85.6% homology. The expression profiles of VfBCCP gene were investigated by employing quantitative real-time RT-PCR method. The results showed that the lower transcription levels of VfBCCP was presented at the early stage of α-eleostearic acid biosynthesis in the middle of July, and rapidly increased during the eleostearic acid accumulation in August, then decreased during accumulation completion period at the end of September. The isolation and expression rule investigation could hold the promise of clarifying VfBCCP function in fatty acid biosynthesis.

Key words: Vernicia fordii, biotin carboxyl carrier protein (BCCP), gene cloning, expression analysis