苎麻生长素结合蛋白ABP1基因cDNA的克隆及表达  

Cloning and Expression of Auxin-binding Proteins 1 Gene in Ramie[Boehmeria nivea(Linn.) Gaud.]

作　　者：黄妤[1] 刘峰[2] 郭清泉[2] 张学文[1] 

机构地区：[1]湖南农业大学生物科学技术学院,湖南长沙410128 [2]湖南农业大学苎麻研究所,湖南长沙410128

出　　处：《作物学报》2008年第8期1358-1365,共8页Acta Agronomica Sinica

基　　金：国家自然科学基金项目(30571186);湖南省自然科学基金项目(08JJ5020)

摘　　要：以苎麻[Boehmeria nivea(Linn.)Gaud.]栽培种湘苎3号为材料,通过简并引物RT-PCR结合RACE技术克隆了苎麻生长素结合蛋白ABP1基因的全长cDNA分子,其序列全长为849bp,编码一段189个氨基酸的推导蛋白质。经基因比对及蛋白质结构分析与已报道的几种植物生长素结合蛋白有高同源性,认为是苎麻生长素结合蛋白基因cDNA,命名为BnABP1。半定量RT-PCR分析结果显示ABP1在苎麻三麻成熟期的茎、叶和芽组织中均有表达,其表达量为芽>叶>茎,相对内标分子18S rRNA的表达量依次为0.755、0.632和0.360。但在根中没有检测到表达,说明该基因更多地表达于细胞生长旺盛的幼嫩组织。Auxin, a kind of phytohormone discovered quite earlier, affects many processes of plant growth. If we identify the receptor interplaying with auxin directly and find out how the signal transfered into the tissue cells, it can help us to learn the mechanism of auxin effection, detecting many processes about plant＇s growth deeply. ABP1 （auxin binding protein 1） was cloned from upland cotton （Gossypium hirsutum Linn.）, capsicum, etc. But there is not any report about the auxin binding protein of ramie. In this paper the ABP1 cDNA core sequence was cloned by PCR with the primers designed using Boehmeria nivea（Linn.） Gaud as material. The identical molecules were cloned through 5＇ and 3＇ RACE and the whole sequence of the cDNA was cloned and sequenced which was a 849 bp molecule and could be translated into putative protein with 189 amino acids. BLAST analysis confirmed that this cDNA sequence shared a high homology with reported ABP gene of plants. It was designated as BnABP1 according to the auxin binding protein gene nomination habit and was submitted to GenBank with an accession number EU195804. In order to investigate the expression and regulation roles of BnABP1 in different tissues of ramie, the non-conservative sequence at 3＇-end of BnABP1 was selected as target and its expression was detected by semi-quantitative RT-PCR with 18S rRNA as internal control. After running the PCR with various cycles the products electrophoresed in agarose gel and the integrating optic density （IOD） of bands was detected with gel analysis software. We take the ratio of IOD as relative expressive quantity. The results indicated the expression of BnABP1 could be found in leaf, stem and bud but not in root in ramie. The expression level in bud, leaf and stem was with the relative content of 0.755, 0.632, and 0.360 respectively compared with that of 18S rRNA. Thus BnABP1 maybe expresses mostly in the tender tissues of plant.

关 键 词：苎麻 生长素结合蛋白 CDNA克隆 表达分析 

分 类 号：S563.1[农业科学—作物学]