华北农学报 ›› 2009, Vol. 24 ›› Issue (5): 26-30. doi: 10.7668/hbnxb.2009.05.006

所属专题: 生物技术

• 论文 • 上一篇    下一篇

家蚕黄酮合酶Ⅰ基因BmFNS Ⅰ的克隆与干涉载体的构建

孔卫青, 杨金宏, 彭云武, 胡必利   

  1. 安康学院陕西省蚕桑重点实验室, 陕西安康 725000
  • 收稿日期:2009-07-09 出版日期:2009-10-28
  • 作者简介:孔卫青(1980-),女,山东成武人,博士,讲师,主要从事家蚕分子遗传学研究。
  • 基金资助:
    陕西省自然科学基金项目(SJ08C108);安康学院专项科研计划资助项目(07AKXY026)

Cloning and Construction of RNAi Vector of Flavone Synthase I Gene, BmFNS I from Silkworm,Bombyx mori

KONG Wei-qing, YANG Jin-hong, PENG Yun-wu, HU Bi-li   

  1. The Key Sericultural Laboratory of Shaanxi, Ankang University, Ankang 725000, China
  • Received:2009-07-09 Published:2009-10-28

摘要: 黄酮类化合物广泛存在于动植物中,具有多种生理活性,黄酮合酶Ⅰ基因可以催化多种黄酮类化合物的生物合成。根据NCBI已登录的其他物种黄酮合酶Ⅰ基因的氨基酸序列,与家蚕基因组和EST数据库进行电子克隆获得家蚕的同源基因BmFNS Ⅰ。克隆测序结果表明该基因长1451 bp,7个外显子。根据基因序列推测其编码蛋白由345个氨基酸构成,179~295间氨基酸为一2-氧戊二酸和Fe(Ⅱ)依赖性的加氧酶家族成员的结构域。RT-PCR检测该基因在本试验所调查的家蚕中肠有高表达。最后将该基因片段亚克隆至具有2个反向T7启动子可以用于体外诱导合成dsRNA的L4440载体。

关键词: 家蚕, BmFNS Ⅰ基因, 克隆, 组织表达

Abstract: Flavone is widely existing in animals and plants.They have various physiological activity and the synthesization of them can be catalyzed by flavone synthase I gene.Using the reported amino acid sequences of other organisms and the scaffold and EST sequences of silkworm,silkworm flavone synthase I gene,BmFNS I,was obtained by electronic clone.The gene is 1451 bp in length and 7 exons in it by cloning and sequencing.The deduced protein contains 345 amino acids and a structural domain of 2-oxoglutarate (2OG) and Fe(II)-dependent oxygenase superfamily located in amino acids 179 and 295.It is shown by RT-PCR experiment that the gene expressed mostly in midgut in all the tissues tested.Then gene was subcloned to L4440,which has two T7 polymerase promoter in opposite orientation to synthesize dsRNA in vitro.

Key words: Bombyx mori, BmFNS I, Clone, Tissue expression

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引用本文

孔卫青, 杨金宏, 彭云武, 胡必利. 家蚕黄酮合酶Ⅰ基因BmFNS Ⅰ的克隆与干涉载体的构建[J]. 华北农学报, 2009, 24(5): 26-30. doi: 10.7668/hbnxb.2009.05.006.

KONG Wei-qing, YANG Jin-hong, PENG Yun-wu, HU Bi-li. Cloning and Construction of RNAi Vector of Flavone Synthase I Gene, BmFNS I from Silkworm,Bombyx mori[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2009, 24(5): 26-30. doi: 10.7668/hbnxb.2009.05.006.

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