抗玉米赤霉烯酮毒素单链抗体-碱性磷酸酶融合蛋白的表达

根据抗玉米赤霉烯酮毒素单链抗体的氨基酸序列及大肠杆菌偏爱密码子,用重叠延伸PCR法合成了单链抗体的重链和轻链,经（Gly4Ser）2Linker连接,获得完整的单链抗体基因ZEN2 scFv。将ZEN2 scFv与碱性磷酸酶（AP）编码序列连接形成融合蛋白基因ZEN2 scFv-AP,构建到pET载体,转化大肠杆菌菌株BL21（DE3）,经IPTG诱导表达及亲和层析纯化,在SDS-PAGE和Western blot分析中均检测到1条分子质量为75 ku的可溶性蛋白条带,ELISA分析证实,细菌表达的ZEN2 scFv-AP融合蛋白具有碱性磷酸酶活性。这一研究结果为建立快速、灵敏、经济的玉米赤霉烯酮毒素的ELISA检测奠定了基础。

Expression of Fusion Protein Containing Alkaline Phosphatase and Single-chain Antibody Specific for Zearalenone Mycotoxin

Based on the amino acid sequence of a single-chain antibody specific for zearalenone mycotoxin and codon usage in E.coli,a variable heavy chain(VH) and a variable light chain(VL) were synthesized by PCR to create a single-chain(scFv) antibody gene,ZEN2,that was linked by a(Gly4Ser)2 linker.The scFv antibody gene,ZEN2,was further ligated into an alkaline phosphatase(AP) sequence to construct a fusion protein,ZEN2 scFv-AP that was cloned into pET vector.E.coli strain BL21(DE3) was transformed with the pET pla...