用二次电泳法研究核酸与蛋白质的相互作用

Reelectrophoreses in different buffer:a new method for DNA or RNA binding assay of proteins

研究蛋白质与核酸的结合常遇到的问题是对蛋白质等电点及可溶度等要求较高，或难以同时处理大量标本。为克服此缺点，将待检蛋白经聚丙烯酰胺凝胶电泳后，通过洗涤去除凝胶中的ＳＤＳ，使蛋白质相对固定于凝胶中，改电泳液为ＴＡＥ或ＴＢＥ，继之用同位素标记寡核苷酸进行二次电泳，通过放射自显影直观地显现出蛋白结合核酸的结果。该法敏感，特异，对蛋白质等电点及可溶性要求低，可同时检测多个样本，值得推广使用。

To overcome some defects in known DNA-protein and RNA-protein binding experi- ments we electrophoresed proteins through SDS-PAGE and then removed the SDS from the gel by washing with salty buffer and TAE. The gel was set again and γ-32P ATP labeled oligonu- cleotides were electrophoresed through the gel in TAE buffer. The binding signals were dtected by autoradiography. Proteins detected remains the specific binding activity after their renatura- tion.