中国水仙叶绿体ATP合成酶基因NtATPF的克隆与序列分析

Cloning and Sequence Analysis of Chloroplast ATP Synthase Gene NtATPFfrom Narcissus tazetta var.Chinensis

摘要: 以多效唑诱导中国水仙的抑制差减杂交文库 (SSH) 获得的cDNA片段为基础, 采用RACE及RT-PCR技术, 分离克隆到1个ATP合成酶CF0B亚基基因NtATPF。此基因包含有1个555bp完整阅读框架 (ORF) , 编码184个氨基酸, 具有1个跨膜区域 (27⁴9) , 分子量为20.809 5kD, 理论等电点为5.69。聚类分析表明:其氨基酸序列与葱的叶绿体ATP合成酶CF0B亚基蛋白氨基酸序列亲缘关系最近。

Abstract: Based on a fragment cDNA from suppression subtractive hybridization (SSH) of Narcissus tazetta var.chinensis induced by paclobutrazol, an ATP Synthase CF0 subunit gene NtATPF was separated and cloned by RACE and RT-PCR.The full length of NtATPFcDNA was 905 bp with a 555 bp ORF, which encoded a protein of184 amino acids and had a transmembrance domain (27-49) with a predicted molecular mass of 20.809 5kD and theoretical isoelectric point of 5.69.Phylogenetic analysis showed that NtATPF had close relationship to ATP Synthase CF0 subunit in Allium fistulosum (AEX93661.1) .