摘要
RNA结合蛋白是基因表达的重要调节因子.RNA蛋白质印迹(Northwesternblot)是近年来国外建立的筛选这类因子的重要方法之一.应用这一方法从肝细胞株HepG2cDNA文库中成功筛选到乙型肝炎病毒表面抗原转录后调节片段互相作用蛋白表达克隆.结果显示:该蛋白与探针结合特异性强,经三轮筛选后,100%克隆为阳性克隆;PCR和EcoRⅠ酶切初步鉴定,编码该蛋白的cDNA长约1kb.
Northwestern Blot has been used for isolation of cDNAs encoding proteins interacting with HBsAg posttranscriptional regulatory element (PRE). One expression cDNA clone which showed specially binding to PRE was isolated from λgt11 cDNA expression library prepared with human hepatoma cell line HepG2. The size of this PRE interacting protein(PIP) clone was identified to be about 1 kb by PCR and digestion with restriction endonuclease Eco RⅠ.The results showed that Northwestern blot with some modifications is a useful method for isolation of expression clones of genes encoding RNA binding proteins.
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
2000年第1期75-78,共4页
Progress In Biochemistry and Biophysics
基金
国家自然科学基金!资助项目 (39670 653)
关键词
RNA-蛋白质印迹
HBSAG
PRE
调节因子
分离
PIP
Northwestern blot, hepatitis B virus surface antigen, posttranscriptional regulatory element ,PRE interacting proteins
