过表达<em>cdc20</em>基因对绒山羊卵母细胞体外成熟的影响

doi:10.7668/hbnxb.2015.06.005

华北农学报 ›› 2015, Vol. 30 ›› Issue (6): 32-36. doi: 10.7668/hbnxb.2015.06.005

所属专题： 畜牧；生物技术；

过表达cdc20基因对绒山羊卵母细胞体外成熟的影响

张金吨¹, 王标¹, 李云霞^1,2, 陈杨林¹, 刘黎明¹, 戴雁峰^1,2, 李喜和^1,2

1. 内蒙古大学生命科学学院, 蒙古高原动物遗传资源研究中心, 内蒙古呼和浩特 010021;
2. 内蒙古赛科星繁育生物技术(集团)股份有限公司, 内蒙古呼和浩特 011517

收稿日期:2015-09-20 出版日期:2015-12-28
通讯作者: 戴雁峰(1963-),男,辽宁海城人,教授,博士,主要从事哺乳动物性腺发育及配子形成的研究;李喜和(1963-),男,内蒙古呼和浩特人,教授,博士,主要从事哺乳动物性控技术的研究.
作者简介:张金吨(1985-),男,内蒙古包头人,在读博士,主要从事哺乳动物生殖生物学及生物技术研究.
基金资助:
国家自然科学基金面上项目(31272445);内蒙古高原动物遗传资源库建立与利用(20121406;20130216)

Effect of Overexpression of cdc20 during Cashmere Goat Oocyte Maturation in vitro

ZHANG Jin-dun¹, WANG Biao¹, LI Yun-xia^1,2, CHEN Yang-lin¹, LIU Li-ming¹, DAI Yan-feng^1,2, LI Xi-he^1,2

1. College of Life Sciences, Inner Mongolia University, Research Center for Animal Genetic Resources of Mongolian Plateau, Huhhot 010021, China;
2. Inner Mongolia Saikexing Reproductive Biotechnology Co., LTD, Huhhot 011517, China

Received:2015-09-20 Published:2015-12-28

摘要/Abstract

摘要： 为研究过表达cdc20 基因对绒山羊卵母细胞体外成熟的影响,构建了Myc-cdc20 载体,在体外转录成cRNA后显微注射到绒山羊卵母细胞中,观察其对绒山羊卵母细胞体外成熟过程的影响.结果表明,在对照组卵母细胞体外培养的过程中,卵母细胞培养到8 h后发生GVBD,18 h后进入MⅠ期,直到24 h后,约75%的绒山羊卵母细胞排出第一极体.卵母细胞在GV期时显微注射gMyc-cdc20 cRNA,培养10 h后,通过Western Blotting的方法检测到了MYC-CDC20融合蛋白的表达.但统计结果显示,试验组卵母细胞各阶段发育时间和成熟率与对照并无显著差异.因此,单独过表达cdc20 基因不影响绒山羊卵母细胞体外成熟过程.

关键词: 过表达, cdc20, 绒山羊卵母细胞, 体外成熟

Abstract: In order to investigate the effect of overexpression of cdc20 during cashmere goat oocyte maturation in vitro ,gMyc- cdc20 vector was constructed and overexpressed in the oocyte by microinjection after transcripted into cRNA in vitro.The results showed that the normal oocytes needed 8 h from GV stage to GVBD stage,and 18 h to MⅠ.Twenty-four hours later,75% of these oocytes extruded first polar body.Then microinjected gMyc- cdc20 cRNA into GV stage oocytes and cultured them in vitro.MYC-CDC20 fusion protein could be detected by Western Blotting after 10 h.However,both of the development time and the ratio of oocyte maturation were not significantly different compared with those of control.It could conclude that overexpression of cdc20 alone had no effect on the oocyte maturation of cashmere goat in vitro.

Key words: Overexpression, cdc20, Cashmere goat oocyte, Maturation in vitro

中图分类号:

S826

张金吨, 王标, 李云霞, 陈杨林, 刘黎明, 戴雁峰, 李喜和. 过表达cdc20基因对绒山羊卵母细胞体外成熟的影响[J]. 华北农学报, 2015, 30(6): 32-36. doi: 10.7668/hbnxb.2015.06.005.

ZHANG Jin-dun, WANG Biao, LI Yun-xia, CHEN Yang-lin, LIU Li-ming, DAI Yan-feng, LI Xi-he. Effect of Overexpression of cdc20 during Cashmere Goat Oocyte Maturation in vitro[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2015, 30(6): 32-36. doi: 10.7668/hbnxb.2015.06.005.

http://www.hbnxb.net/CN/Y2015/V30/I6/32

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Effect of Overexpression of cdc20 during Cashmere Goat Oocyte Maturation in vitro

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