摘要
根据国外已发表的鹅细小病毒(GPV)A株基因组核苷酸序列,设计并合成了一对用于GPV主要结构蛋白(VP2-VP3)基因表达的引物。利用合成的引物,扩增并鉴定了GPV中国长春株(GPVCC株)的VP2-VP3基因。将扩增的目的基因插入到原核表达载体pET28(a)的多克隆位点,构建了表达GPVVP2-VP3基因的原核载体pEGVP1。重组表达载体质粒转化BL21宿主菌,经IPTG诱导,SDS-PAGE检测到大小分别为75000和58000的表达蛋白带。Westernblot分析表明,表达产物具有很好的特异性。
According to nucleotide suquence of GPV A strain,a pairs of primers were designed and synthesized,and mutated the VP2ACG codon to ATG one.The complete VP2-VP3(the major structural protein)genes of a Chinese isolate stain(GPV CC)was amplified by PCR,and obtained a DNA fragment with the sizes of1.8kb.The PCR prod-uct was digested with Bam HI and the resulting fragment was inserted into the Bam HI site of the pET28(a)under the downstream sequences.E.coli component host BL21was transformed with the recombinant.The recombinant bac-teria could express the VP2-VP3gene after induced by IPTG,and the recombinant protein were analyzed on12%SDS-PAGE and stained with comass brilliant blue or had been detected by immunblotting analysis with anti-GPV serum.
出处
《生物技术通讯》
CAS
2002年第5期331-333,共3页
Letters in Biotechnology
基金
国家自然科学基金项目(39870557)
教育部和总后勤部回国人员启动基金项目(98H026)
