CAJ | 학술논문

为获得针对番鸭细小病毒VP3蛋白的多克隆抗体,根据已发表的该病毒基因序列设计一对引物,利用PCR扩增出VP3基因,将其克隆到原核表达载体pET-32a,转化感受态细胞BL21(DE3)。经IPTG诱导后,SDS-PAGE检测重组蛋白。将重组蛋白切胶免疫BALB/c小鼠,制备抗番鸭细小病毒部分结构蛋白的多克隆抗体。结果显示成功获得VP3基因,构建的原核表达重组质粒成功表达预期的重组蛋白,免疫小鼠获得的多克隆抗体能与番鸭细小病毒及VP3蛋白反应。表明制备的多克隆抗体可用于MDPV的检测,并为进一步研究MDPV奠定了基础。
위획득침대번압세소병독VP3단백적다극륭항체,근거이발표적해병독기인서렬설계일대인물,이용PCR확증출VP3기인,장기극륭도원핵표체재체pET-32a,전화감수태세포BL21(DE3)。경IPTG유도후,SDS-PAGE검측중조단백。장중조단백절효면역BALB/c소서,제비항번압세소병독부분결구단백적다극륭항체。결과현시성공획득VP3기인,구건적원핵표체중조질립성공표체예기적중조단백,면역소서획득적다극륭항체능여번압세소병독급VP3단백반응。표명제비적다극륭항체가용우MDPV적검측,병위진일보연구MDPV전정료기출。
To prepare po1yc1ona1 antibody of VP3 protein of Muscovy duck parvovirus( MDPV) , one pair of specific primers was designed according to the pub1ished genome sequences of MDPV to amp1ify VP3 gene by PCR. The amp1ified fragment was c1oned into prokaryotic expression vector pET-32a and the identified recombinant p1asmid was transformed into Escherichia. coli BL21(DE3). The protein was expressed fo11owing IPTG induction and detected by the SDS-PAGE. Then the recombinant protein was used to immunize Ba1b/c mice to prepare the po1yc1ona1 antibodies . The resu1ts showed that the VP3 gene was successfu11y expressed in prokaryotic expression vector, and the po1yc1ona1 antibody cou1d react with VP3 proteins and MDPV, suggesting the po1yc1ona1 antibody can be used for the identify of MDPV.