花生野生近缘种生物素羧基载体蛋白基因的克隆与结构分析

Cloning and Structural Analysis of Biotin Carboxyl Carrier Protein Genes in Wild Relatives of Arachis hypogaea

根据栽培花生鲁花14生物素羧基载体蛋白基因accB1和accB2cDNA序列设计基因特异引物,克隆了花生野生近缘种Arachis duranensis、A.rigonii、A.batizocoi和A.hoehnei的accB1和accB2cDNA序列。序列分析表明,accB1和accB2在栽培花生和野生近缘种中高度保守,氨基酸序列一致性分别高于98.6%和97.5%。A基因组A.duranensis、A.rigonii和B基因组A.batizocoi、A.hoehnei的accB1和accB2的氨基酸序列存在差异,但与基因组来源无关。花生野生近缘种A.duranensis、A.rigonii、A.batizocoi和A.hoehnei accB2基因组序列分析表明,栽培花生和野生近缘种的accB2的基因结构相同。A基因组的A.duranensis与B基因组的A.batizocoi和A.hoehnei的accB2基因组DNA核苷酸序列一致性分别为98.9%,98.8%,而A基因组的A.rigonii与A.duranensis、A.batizocoi和A.hoehnei的序列一致性分别为93.5%,93.6%,93.4%。研究表明,生物素羧基载体蛋白基因在栽培花生及野生近缘种中非常保守。

Gene-specific primers of accB1,accB2 were designed based on cDNA sequences of cultivated peanut Luhua-14 biotin carboxyl carrier protein（BCCP） genes.The accB1 and accB2 cDNA sequences of peanut wild relatives,A.duranensis,A.rigonii,A.batizocoi and A.hoehnei were isolated by PCR using leaf cDNA as templates.accB1 and accB2 were highly conserved in peanut and its wild relatives and amino acid sequences of accB1 and accB2 showed 98.6% and 97.5% identity.The intron-exon structure of the accB2 of peanut wild relatives,A.duranensis,A.rigonii,A.batizoco,A.hoehnei,was identical with that of Luhua-14.Genomic DNA sequence of A.duranensis shared 98.9%,98.8% nucleotide sequence identity with A.batizocoi and A.hoehnei,while genomic DNA sequence of A.rigonii showed 93.5%,93.6%,93.4% nucleotide sequence identity with that of A.duranensis,A.batizocoi,A.hoehnei,respectively.The results indicated that accB1 and accB2 were highly conserved in peanut and its wild relatives.