花生 mtACP3 基因的克隆与表达分析

doi:10.11924/j.issn.1000-6850.casb16100110

中国农学通报 ›› 2017, Vol. 33 ›› Issue (20): 35-42.doi: 10.11924/j.issn.1000-6850.casb16100110

所属专题：生物技术；油料作物

花生 mtACP3 基因的克隆与表达分析

迟晓元,陈娜,王通,王冕,陈明娜,潘丽娟,郝翠翠,杨珍,梁成伟,禹山林

山东省花生研究所,山东省花生研究所,山东省花生研究所,山东省花生研究所,青岛市李沧区万年泉路126号,青岛市李沧区万年泉路126号,青岛科技大学,青岛市李沧区万年泉路126号,青岛科技大学,青岛市李沧区万年泉路126号

收稿日期:2016-10-27 修回日期:2017-06-29 接受日期:2016-12-23 出版日期:2017-07-18 发布日期:2017-07-18
通讯作者: 禹山林
基金资助:
2014 年国家万人计划“青年拔尖人才”；国家花生产业技术体系“良种扩繁与种子生产岗位”(CARS-14)；山东省三院联合基金“花生kennedy 途径关键酶基因的分离与功能研究”(ZR2014YL012)，“花生连作根腐病的主效微生物挖掘及其调控因子分析”(ZR2014YL011)；国家自然基金“参与花生脂肪酸合成代谢的关键酶基因的克隆与功能研究”(31000728)，“花生iso-Ara h3 基因在种子抗黄曲霉中的功能研究”(31200211)；山东省农业科学院青年英才培养计划“访学研修”。

Cloning and Expression Analysis of mtACP3 Genes in Peanut

Received:2016-10-27 Revised:2017-06-29 Accepted:2016-12-23 Online:2017-07-18 Published:2017-07-18

摘要/Abstract

摘要： 旨在探究酰基载体蛋白在植物脂肪酸合成途径中的作用机制，本研究采用传统PCR方法从花生中克隆得到了一个线粒体型基因，命名为AhmtACP3。并采用荧光定量PCR技术分析了基因的表达特性。该基因全长为859 bp，开放阅读框ORF为390 bp，开放阅读框对应的基因组序列为543 bp，由2 个外显子和1 个内含子组成，该基因编码129 个氨基酸，理论分子量为14.5345 kD，理论等电点为5.22，可能定位于线粒体上。系统发育分析表明，花生线粒体型ACP 蛋白分成2 个分支：AhmtACP1 和AhmtACP2 有较近的亲缘关系，与AhmtACP3 亲缘关系较远。荧光定量PCR结果显示，AhmtACP3 基因在花中的表达量明显高于其他组织，其次是子叶和根。AhmtACP3 基因在种子发育过程中的表达量呈现下降趋势。

关键词: 酿造高粱, 酿造高粱, 肥料, 高效利用

Abstract: To investigate the mechanism of acyl carrier protein (ACPs) in the pathway of plant fatty acid synthesis, one ACP gene (AhmtACP3) was isolated from peanut by traditional PCR. The expression characteristics of the gene were analyzed by quantitative real- time RT- PCR (qRT- PCR). The length of AhmtACP3 gene was 859 bp. The length of open reading flame (ORF) was 390 bp, and its genomic sequence was 543 bp with 2 exons and 1 intron. It encoded a protein of 129 deduced amino acid residues. The molecular weight was 14.5345 kD and its isoelectric point was 5.22. The deduced protein was predicted to locate in mitochondria. Phylogenetic analysis indicated that peanut mitochondrial ACPs fell into two clusters. AhmtACP1 was grouped with AhmtACP2, which was distinct from AhmtACP3. qRT- PCR indicated that AhmtACP3 transcripts were more abundant in flowers than in any other tissues tested, followed by cotyledons and roots. The expression levels ofAhmtACP3 were the highest at the initial stage of seed development, but gradually decreased in abundance during the later stages.

迟晓元,陈娜,王通,王冕,陈明娜,潘丽娟,郝翠翠,杨珍,梁成伟,禹山林. 花生 mtACP3 基因的克隆与表达分析[J]. 中国农学通报, 2017, 33(20): 35-42.

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花生 mtACP3 基因的克隆与表达分析

Cloning and Expression Analysis of mtACP3 Genes in Peanut

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