华北农学报 ›› 2018, Vol. 33 ›› Issue (1): 96-101. doi: 10.7668/hbnxb.2018.01.015

所属专题: 油料作物 生物技术

• 论文 • 上一篇    下一篇

甘蓝型油菜BnACP5基因克隆及表达分析

贺慧, 虢慧, 官春云   

  1. 国家油料改良中心湖南分中心, 湖南 长沙 410128
  • 收稿日期:2017-11-22 出版日期:2018-02-28
  • 通讯作者: 官春云(1938-),男,湖南长沙人,教授,博士生导师,主要从事油菜育种研究。
  • 作者简介:贺慧(1991-),男,湖南株洲人,在读硕士,主要从事油菜分子育种研究。
  • 基金资助:
    国家"973"计划项目(2015CB150206);"双季稻区油菜早熟品种选育和机械化生产技术" (湘财农指(2016)114号)

Cloning and Expression Analysis of BnACP5 in Brassica napus

HE Hui, GUO Hui, GUAN Chunyun   

  1. National Oilseed Crops Improvement Cent in Hunan, Changsha 410128, China
  • Received:2017-11-22 Published:2018-02-28

摘要: ACP5基因是参与植物体中脂肪酸代谢的重要基因之一,在甘蓝型油菜中该基因还未被深入研究。为了弄清楚BnACP5基因在甘蓝型油菜中的螺旋结构及功能,以湘油15为试材,采用同源克隆法得到一个与脂肪酸合成相关的基因BnACP5BnACP5基因CDs序列长417 bp,共编码138个氨基酸。生物信息分析表明,BnACP5蛋白的相对分子质量15.25 ku,理论等电点(pI)为5.94,其不稳定参数为43.03,属于不稳定蛋白;其总疏水平均系数(GRAVY)为-0.148,是一种亲水性蛋白;不含跨膜结构,也不含信号肽序列;二级结构中含有59.42%的α螺旋、23.19%无规则卷曲、14.49%延伸链和2.90%β转角;其中18个可能是磷酸化位点;该蛋白位被定位在线粒体内;ACP氨基酸序列比对表明,植物ACP磷酸泛酰巯基乙胺结合位点周围的氨基酸序列高度保守;UPGMA聚类分析表明,BnACP5与白菜型油菜(XP_009111933.1)亲缘关系最近。荧光定量PCR结果显示,BnACP5基因的表达量在甘蓝型油菜种子发育的不同阶段呈一定规律变化,在授粉后15 d达到峰值,20 d呈下降趋势,25~35 d呈上升趋势,此后迅速下降,45 d表达量降到最低水平。由此分析得出,该基因可能与种子脂肪酸的合成有关。

关键词: 甘蓝型油菜, BnACP5, 基因克隆, 荧光定量PCR

Abstract: ACP5 gene is one of the important genes involved in the metabolism of fatty acids in plants,the gene has not been studied in depth,in Brassica napus.In order to clarify the structure and function of BnACP5 gene in Brassica napus.A gene related to fatty acid synthesis, BnACP5,was obtained by homologous cloning,Xiangyou 15 was used as test material.We cloned a full length CDs sequence that was 417 bp and encoded 138 amino acid residues.Bioinformaties analyses showed that BnACP5 was an unstable protein with a molecular weight of 15.25 ku,a theoretical isoelectric point of 5.94,instability index of 43.03 and a hydrophilic protein with the GRAVY of -0.148, and 18 of them might be phosphorylation sites,but signal peptide sequence and transmembrance domain were not found.The secondry structure of BnACP5 protein contained 59.42% of alpha helix,23.19% of random coil,14.49% of extended strand and 2.90% of beta turn.BnACP5 protein was tested to be a typical acyl carrier protein localized in mitochondria.ACP amino acid sequence alignment showed that the amino acid sequence around the plant ACP phosphopantetheine binding site was highly conserved.UPGMA tree analysis indicated that BnACP5 protein was closely related to the ACP5 in B. rapa. RT-PCR results showed that BnACP5 expressed in oilseeds specially.qRT-PCR results indicated that the expression of BnACP5 reached the highest level at the 15th day after flowering,and decreased at the 20th day,and increased from 25 to 35 days, after rapid decline,and 45 d expression was reduced to the lowest level.These results demonstrated that BnACP5 might be associated with the synthesis of fatty acids in oilseeds.

Key words: Brassica napus, BnACP5, Gene cloning, qRT-PCR

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引用本文

贺慧, 虢慧, 官春云. 甘蓝型油菜BnACP5基因克隆及表达分析[J]. 华北农学报, 2018, 33(1): 96-101. doi: 10.7668/hbnxb.2018.01.015.

HE Hui, GUO Hui, GUAN Chunyun. Cloning and Expression Analysis of BnACP5 in Brassica napus[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2018, 33(1): 96-101. doi: 10.7668/hbnxb.2018.01.015.

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