摘要
SDHB（succinate dehydrogenase complex，subunit B）基因可能介导呼吸链生物功能和调控细胞生长。采用PCR技术扩增出SDHB基因，并将其连接到pGEX-4T-1原核表达载体中，经酶切及测序鉴定后，转化BL21细菌，并用IPTG诱导表达融合蛋白，谷胱甘肽琼脂糖珠亲和纯化，将纯化的融合蛋白免疫新西兰大白兔制备多克隆抗体，并用Western blot检测抗体。获得了SDHB原核表达重组融合蛋白及高效价的特异性兔抗SDHB多克隆抗体，为SDHB进一步的功能研究奠定了基础。
The SDHB（ succinate dehydrogenase complex,subunit B）gene might play a role in mediating the biological function of respiration chain and cellular growth. SDHB gene was amplified by method of PCR technique, then was cloned into pGEX-4T-1 vector and identified with restriction enzyme and sequencing. The recombinant expression plasmid containing SDHB gene was transformed into BL21 and fusion protein was induced by IPTG. The New Zealand white rab- bits was immuned with the purity protein obtained by treating the lysates with immobilized Glutathione Sepharose to prepare antibody. The antibody titer and specification was identified by Western Blot. We have successfully obtained GST fusion protein of SDHB, got high sensitivity and specificity anti-SDHB polyclonal antibody. That lays a solid foundation for the further studies of SDHB function.
出处
《激光生物学报》
CAS
CSCD
2009年第3期365-369,共5页
Acta Laser Biology Sinica
基金
国家自然科学基金项目(30671053,30871340)
