牡丹CTR1基因家族基因片段及其cDNA 3′-末端的克隆与序列分析

Cloning and Sequence Analysis of CTR1 Gene Family Sequences and Its 3′cDNA Ends from Tree Peony

以牡丹品种‘洛阳红’(Paeonia suffruticosa‘Luo Yang Hong’)花瓣为材料,用CTAB法提取总RNA,根据已报道的CTR1(Constitutive Triple Response 1)蛋白的保守氨基酸序列设计简并引物,通过RT-PCR扩增得到3条牡丹CTR1基因同源片段。利用其已知中间序列,通过3′快速扩增cDNA末端技术(3′RACE)及序列拼接,最终得到了这3个基因片段除5′末端外的全部cDNA序列,分别命名为PsCTR1、PsCTR2和PsCTR3。分析结果表明,由PsCTR1、PsCTR2和PsCTR3基因翻译的氨基酸片段与拟南芥和番茄的CTR1蛋白氨基酸序列的同源性均较高,分别为88%和85%;61%和61%以及70%和69%。

Total RNA was extracted from petals of ＇Luoyang Hong＇ tree peony（Paeonia suffruticosa）with CTAB method,and two pairs of degenerated primers were designed based on the reported conserved amino acid sequence of Constitutive Triple Response 1（CTR1）.Three cDNA fragments were amplified by RT-PCR,and then their sequences except the 5′cDNA end were obtained by 3′RACE and sequence spliced.They are named PsCTR1,PsCTR2 and PsCTR3.Sequence analysis indicated that the predicated amino acid residues which encodes shared more than 60% homology with CTR1 of Arabidopsis,tomato and so on.