酵母双杂交方法对牛CMYA1中Xin Repeat结构域的鉴定

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

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。

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.