家蚕,线粒体,细胞色素C,细胞凋亡," /> 家蚕,线粒体,细胞色素C,细胞凋亡,"/> Bombyx mori,mitochondrial,cytochrome c,apoptosis,"/> <font face="Verdana">家蚕细胞色素C基因的克隆及其蛋白在家蚕凋亡细胞中的释放</font>

中国农业科学 ›› 2009, Vol. 42 ›› Issue (7): 2546-2551 .doi: 10.3864/j.issn.0578-1752.2009.07.036

• 畜牧·资源昆虫 • 上一篇    下一篇

家蚕细胞色素C基因的克隆及其蛋白在家蚕凋亡细胞中的释放

潘敏慧,陈 默,黄淑静,于子舒,成传刚,鲁 成   

  1. (西南大学蚕学与系统生物学研究所,农业部蚕桑学重点开放实验室)
  • 收稿日期:2008-08-28 修回日期:2008-09-10 出版日期:2009-07-10 发布日期:2009-07-10
  • 通讯作者: 鲁成

Cloning and Protein Release of Cytochrome C in the Apoptosis Cells of Silkworm, Bombyx mori#br#

PAN Min-hui, CHEN Mo, HUANG Shu-jing, YU Zi-shu, CHENG Chuan-gang, LU Cheng#br#   

  1. (西南大学蚕学与系统生物学研究所,农业部蚕桑学重点开放实验室)
  • Received:2008-08-28 Revised:2008-09-10 Online:2009-07-10 Published:2009-07-10
  • Contact: LU Cheng

摘要:

【目的】细胞色素C是线粒体凋亡路径上的关键因子,通过家蚕细胞色素C基因的克隆和其蛋白在家蚕凋亡细胞中的释放研究,为家蚕细胞凋亡的研究奠定基础。【方法】利用生物信息学和分子生物学方法克隆家蚕细胞色素C基因,通过紫外线照射诱导家蚕细胞凋亡,应用流式细胞仪和Western-blotting检测家蚕凋亡细胞的线粒体膜电位变化和细胞色素C的释放。【结果】在家蚕中克隆了一个含有细胞色素C保守结构域的同源基因,该基因cDNA长570 bp,有3个外显子,开放阅读框(open reading frame,ORF)长324 bp,编码108个氨基酸,存在参与细胞凋亡时与下游凋亡蛋白酶激活因子(Apaf-1)相互作用的保守关键位点;紫外线诱导家蚕胚胎细胞BmE-SWU1凋亡后12 h,细胞线粒体跨膜电位明显下降,同时检测到了细胞色素C由线粒体向细胞质释放。【结论】在家蚕细胞凋亡中可能存在细胞色素C由线粒体释放的路径。

关键词: 家蚕')">家蚕, 线粒体, 细胞色素C, 细胞凋亡

Abstract:

【Objective】 Cytochrome c plays a key role in the pathway of mitochondria apoptosis. This study on the cloning and protein release of cytochrome c in the apoptosis cells of silkworm can lay a foundation for the apoptosis research of Bombyx mori. 【Method】 Cytochrome c gene was cloned by bioinformation and molecular biology method. Apoptosis of silkworm cells was induced by UVC. The change of the mitochondrial membrane potential and the release of cytochrome c in apoptosis cells of silkworm were detected by flow cytometry and western-blotting, respectively. 【Result】 The cytochrome c gene was cloned in silkworm (Bombyx mori). The gene contains 3 exons and the length of its cDNA is 570 bp and ORF is 324 bp. The deduced protein has 108 aa and has conservative points for cytochrome c and Apaf-1 interaction related to apoptosis in silkworm. After UVC treated BmE-SWU1 cells for 12 h, the mitochondrial membrane potential decreased and cytochrome c was released from mitochondria to cytoplasm. 【Conclusion】 The cytochrome c may release from mitochondria to cytoplasm in the apoptosis cells in silkworm.

Key words: Bombyx mori')">Bombyx mori, mitochondrial, cytochrome c, apoptosis