水牛睾丸曲精细管蛋白质组双向电泳方法的建立及优化

Establishment and Optimization of Two-Dimensional Electrophoresis for Proteome of Buffalo Testicular Seminiferous Tubules

建立并优化了沼泽型水牛睾丸曲精细管蛋白质分离的双向电泳体系,为后续水牛睾丸曲精细管蛋白质表达谱的鉴定和研究奠定基础。比较不同IPG胶条(线性与非线性)、胶条pH范围和蛋白质上样量这三个参数对双向电泳结果的影响。结果显示,采用上样量350μg的曲精细管总蛋白、24 cm且pH值4～7的线性IPG胶条能够更好地分离水牛睾丸曲精细管蛋白质,获得质量较好且分辨率较高的双向电泳图谱,得到大约486个蛋白点。双向凝胶电泳技术能够对水牛睾丸曲精细管蛋白质进行有效分离,并通过对双向电泳体系的优化可获得蛋白质点清晰且分辨率更高的双向电泳图谱。

A two-dimensional electrophoresis system for separating proteins of swamp buffalo testicular seminiferous tubules was established and optimized,which laid a good foundation for further identification and research of protein expression profiling in the testicular seminiferous tubules of buffalo.The effects of different IPG strips(linear or nonlinear),different pH range of strips and protein sample loading quantity on the results of two-dimensional electrophoresis were compared.The results showed that using 350μg total protein of seminiferous tubules and 24 cm linear IPG strip with pH 4~7 could better separate the protein of buffalo testicular seminiferous tubules,obtain a two-dimensional electrophoresis map with good quality and high resolution,and obtain about 486 protein spots.The two-dimensional gel electrophoresis technology could effectively separate the protein of buffalo testicular seminiferous tubules,and by two-dimensional electrophoresis system optimization,two dimensional electrophoresis map with clear protein spots and higher resolution also could be obtained.