根据茄科植物花色苷生物合成相关基因的资料,设计简并引物,通过RT-PCR的方法从马铃薯(Solanum tuberosum cv.Chieftain)紫色芽的cDNA中克隆到类黄酮-3-O-葡萄糖基化酶基因(3GT)的全长cDNA.序列分析表明,这个3GT基因编码的多肽为448个氨基酸残基,在氨基酸水平上与茄科的矮牵牛和茄子的一致性最高,均为7%,多重比较和系统发育分析均表明,该基因为3GT家族中的一个新成员.用半定量RT-PCR进行了表达分析表明,3GT在根、叶和块茎中的表达量远高于茎和花;此外,3GT在Chieftain的红色愈伤组织中表达而在非红色愈伤组织中不表达,可见,3GT的表达与花色苷的积累是相关的.
The degenerate primers were designed based on the published information of sequence of Solanaceae plants. The cDNA clones encoding UDP2glucose : flavonoid 32O2glucosyltransferase ( 3GT) have been isolated from Solanum tuberosum cv. Chieftain with the degenerate primers by RT2PCR. The cDNA clone of 3GT encodes a predicted polypeptides of 448 amino acids,and the deduce protein showed 76 % highest identity with 3GT of Petunia hybrida and Solanum melongena,and the multiple alignment and phylogenetic analysis demonstrated that it belong to a new member of the corresponding 3GT2family enzymes. The mRNA expression analysis demonstrated that the accumulation of 3GT tran2 scripts in root,leaf and tuber was much more than that in stem and flower. The accumulation of 3GT transcripts in red2 pigmented callus was much more than that in non2pigmented callus of S. tuberosum cv. Chieftain. The expression of 3GT gene followed by anthocyanin accumulation.
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