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园艺学报 ›› 2012, Vol. 39 ›› Issue (8): 1567-.

• 研究报告 • 上一篇    下一篇

结球甘蓝花粉膜联蛋白基因的克隆及其表达分析

 孙梓健, 许俊强, 宋明*, 韦静宜, 汤青林, 王志敏, 王小佳   

  1. (西南大学园艺园林学院,重庆市蔬菜学重点试验室,南方山地园艺学教育部重点试验室,重庆 400715)
  • 出版日期:2012-08-25 发布日期:2012-08-25

Molecular Cloning and Expression Analysis of Annexin Gene from Cabbage

SUN  Zi-Jian, XU  Jun-Qiang, SONG  Ming-*, WEI  Jing-Yi, TANG  Qing-Lin, WANG  Zhi-Min, WANG  Xiao-Jia   

  1. (College of Horticulture and Landscape Architecture,Southwest University;Chongqing Key Laboratory of Olericulture; Key Laboratory of Horticulture Science for Southern Mountainous Regions,Ministry of Education,Chongqing 400715, China)
  • Online:2012-08-25 Published:2012-08-25

摘要: 对结球甘蓝(Brassica oleracea L. var. capitata L.)花粉总蛋白双向电泳及差异蛋白质谱分析, 发现膜联蛋白在花粉萌发后较萌发前表达下调。通过同源扩增和RACE 等方法首次在结球甘蓝花粉中扩增得到BoAnnexin2 基因的cDNA 序列,该基因cDNA 全长1 157 bp,开放阅读框为951 bp,编码316 个氨基酸残基,预测分子量为36.02 kD,等电点6.33。BoAnnexin2 编码蛋白C 端有4 个膜联蛋白重复序列,共2 个Ⅱ型钙结合区域,在第4 个钙结合区位点包含GXXXGXS(T)/DXXG 基序。实时荧光定量试验表明,BoAnnexin2 在甘蓝成熟未萌发花粉中的表达量是萌发45 min 后表达量的8 倍,表明BoAnnexin2 在甘蓝花粉萌发起始阶段起到重要调控作用。

关键词: 结球甘蓝, 花粉, 膜联蛋白

Abstract: Plant annexins constitute a multigene family having suggested roles in a variety of cellular processes. We have isolated and characterized an annexin cDNA of cabbage(Brassica oleracea L. var. capitata L.)gene(BoAnnexin2)encoding annexin proteins using RT-PCR and RACE-PCR after 2-D electrophoresis of cabbage pollen total proteins,which found that annexin protein was downregulated when pollens were germinated. The BoAnnexin2 gene was 1 157 bps,which contained 951 bps open reading frame. The predicted molecular mass of BoAnnexin2 is 36.02 kD with acidic pI of 6.33. At the amino acid level,BoAnnexin2 contains two typeⅡCa2+ binding domain repeats,there was a GXXXGXS (T)/DXXG motif in the fourth domain. The quantitative RT-PCR revealed that the BoAnnexin2 gene expression in mature pollen is eight times higher than that in germinated pollen. These results indicate that BoAnnexin2 gene may play an important role in cabbage pollen germination.

Key words: cabbage, pollen, annexin

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