摘要
根据旋毛虫新生幼虫期特异性基因pBK-CMV-N10序列设计引物,利用PCR技术将基因N10的信号肽去除后,用T/A法克隆到pMD-18T载体,转化至大肠杆菌NovaBlue,经鉴定及序列测定,结果显示成功克隆到N10基因。将重组质粒pMD-18T-N10进行酶切后连接到原核表达载体pET-28a中,重组质粒经鉴定后转化大肠杆菌BL21star(DE3),用IPTG诱导表达出与理论相符的融合蛋白,相对分子质量为38700,诱导4h的表达量占菌体总蛋白量的15%。从N10重组蛋白提取可溶性融合蛋白,对其生物学特性初步鉴定结果表明,具有类似脱氧核糖核酸酶的活性。
Gene N10 was isolated from pBK-CMV-N10 of the newborn larvae cDNA library of T. spiralis by PCR. The positive clones were sequenced and this gene was cloned into pET28a expression vector. The recombined transformant was induced and the recombinant protein of 38 700 was successfully expressed. The DNase Ⅱ activity of the recombinant protein was also identified.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2006年第3期284-287,共4页
Chinese Journal of Veterinary Science
基金
国家自然科学基金资助项目(NSFC30170709
30328020)
中法先进研究计划项目(PRABT03-02)
