酵母双杂交方法对牛CMYA1中Xin Repeat结构域的鉴定

天津农学院学报 ›› 2017, Vol. 24 ›› Issue (1): 28-33.

酵母双杂交方法对牛CMYA1中Xin Repeat结构域的鉴定

王婷, 岳英伟, 杨淑萍, 靳聪飞, 辛向博, 张晓娟, 郭宏^通信作者

天津农学院动物科学与动物医学学院,天津 300384

收稿日期:2016-05-05 出版日期:2017-03-31 发布日期:2019-10-15
通讯作者: 郭宏（1964-）,男,内蒙古通辽人,教授,博士,研究方向为分子遗传与育种。E-mail：guohong64@163.com。
作者简介:王婷（1985-）,女,天津市人,硕士在读,研究方向为临床兽医学。E-mail：wangtingdbnd@126.com。
基金资助:
天津市自然科学基金项目“牛CMYA家族基因结构与功能研究”（11JCZDJC17600）

Identification of Xin Repeat Domain of CMYA1 in Bovine by Yeast Two-Hybrid

WANG Ting, YUE Ying-wei, YANG Shu-ping, JIN Cong-fei, XIN Xiang-bo, ZHANG Xiao-juan, GUO Hong^{Corresponding Author}

College of Animal Science and Veterinary Medicine, Tianjin Agricultural University, Tianjin 300384, China

Received:2016-05-05 Online:2017-03-31 Published:2019-10-15

摘要/Abstract

摘要： CMYA1是肌肉生长发育相关基因,其表达产物包含若干个由16个氨基酸构成的重复序列,该重复序列被命名为Xin Repeat。为确定CMYA1的蛋白结合核心结构域,本研究利用酵母双杂交方法,以牛CMYA1基因的表达产物中含有Xin Repeat的区域作为预测结构域,通过对该区域的不同区段进行克隆建立酵母诱饵表达载体,并转化酵母菌,再与CMYA1的互作蛋白EIF3G和RPL35A进行酵母双杂交。结果表明,共获得了6个不同区段的CMYA1片段,并确定了4个Xin Repeat序列是蛋白结合结构域的最小功能单位。本研究结果有助于揭示CMYA1结构及Xin Repeat的功能,为进一步研究牛肌肉生长发育的分子机制提供理论依据。

关键词: CMYA1, Xin Repeat, 结构域, 酵母双杂交

Abstract: CMYA1 is a muscle development related gene which expression product contains a number of repeat sequences composed of 16 amino acids, named Xin Repeat. To determine the CMYA1 domain of protein interaction, this study used Xin Repeats as a predicted domain of CMYA1. According to different number of Xin Repeats, we divided CMYA1 into different fragments and cloned into expression vectors and transformed into Yeast strain.As result, we obtained 6 fragments of CMYA1 fusion vectors for Yeast two-Hybrid as baits. The minimal unit of protein interacted domain was four Xin Repeats. This study revealed the structure of CMYA1 and the function of Xin Repeats, and provided a theoretical basis of molecular mechanism on bovine muscle growth and development.

Key words: CMYA1, Xin Repeat, domain, Yeast two-Hybrid

中图分类号:

S813.1

王婷, 岳英伟, 杨淑萍, 靳聪飞, 辛向博, 张晓娟, 郭宏. 酵母双杂交方法对牛CMYA1中Xin Repeat结构域的鉴定[J]. 天津农学院学报, 2017, 24(1): 28-33.

WANG Ting, YUE Ying-wei, YANG Shu-ping, JIN Cong-fei, XIN Xiang-bo, ZHANG Xiao-juan, GUO Hong. Identification of Xin Repeat Domain of CMYA1 in Bovine by Yeast Two-Hybrid[J]. Journal of Tianjin Agricultural University, 2017, 24(1): 28-33.

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