猪圆环病毒4型Cap蛋白的原核表达及其多克隆抗体的制备OA
Prokaryotic Expression of the Cap Protein of Porcine Circovirus Type 4 and Preparation of Its Polyclonal Antibody
为获得猪圆环病毒 4 型(porcine circovirus type 4,PCV4)Cap 蛋白及其多克隆抗体,根据 NCBI 公布的 PCV4 HNU-AHG1-2019 毒株基因序列,通过密码子优化合成缺失核定位信号(nuclear localization signal,NLS)的Cap基因.将优化后的基因克隆至pET-28a载体中,成功构建重组表达质粒pET28a-Cap,并转化至BL21(DE3)感受态细胞进行诱导表达.经条件优化发现,在 30℃、0.6 mmol/L IPTG诱导下,约 25 kDa 的 Cap 蛋白以可溶性形式高效表达.纯化后 Cap 蛋白免疫BALB/c小鼠获得的多克隆抗体,抗体效价可达 1∶512 000,Western blot与间接免疫荧光(IFA)实验结果进一步证实,该抗体能够特异性识别 PCV4 Cap蛋白.
In order to prepare polyclonal antibody against Cap protein of porcine circovirus type 4(PCV4),the Cap deleted nuclear location signal(NLS)gene of PCV4 HNU-AHG1-2019 was synthesized based on the codon bias of E.coli and cloned into prokaryotic expression plasmid pET-28a.After verification by enzyme digesting and sequencing,the recombinant positive plasmid pET-28a-Cap was transformed into E.coli BL21(DE3)com-petent cells for induced expression.Through condition optimization,it was found that under the induction of 30℃and 0.6 mmol/L IPTG,approximately 25 kDa of Cap protein was efficiently expressed in soluble form.The purified Cap protein was immunized in BALB/c mice,and the obtained polyclonal antibody had an antibody titer of up to 1∶512 000.The results of Western blot and indirect immunofluorescence(IFA)further confirmed that this antibody could specifically recognize the PCV4 Cap protein.
CAI Yingying;HAN Xueying;DENG Jiaxin;BI Zhen;YAO Lunguang;LIU Yangkun
@@@1.a.School of Life Science||b.Henan Animal and Poultry Health Products Engineering Technology Research Center,Nanyang Normal University,Nanyang Henan 473061,China||2.Department of Modern Service Management,Nanyang Vocational College of Science and Technology,Nanyang Henan 474150,China@@@1.a.School of Life Science||b.Henan Animal and Poultry Health Products Engineering Technology Research Center,Nanyang Normal University,Nanyang Henan 473061,China||2.Department of Modern Service Management,Nanyang Vocational College of Science and Technology,Nanyang Henan 474150,China@@@1.a.School of Life Science||b.Henan Animal and Poultry Health Products Engineering Technology Research Center,Nanyang Normal University,Nanyang Henan 473061,China||2.Department of Modern Service Management,Nanyang Vocational College of Science and Technology,Nanyang Henan 474150,China@@@1.a.School of Life Science||b.Henan Animal and Poultry Health Products Engineering Technology Research Center,Nanyang Normal University,Nanyang Henan 473061,China||2.Department of Modern Service Management,Nanyang Vocational College of Science and Technology,Nanyang Henan 474150,China@@@1.a.School of Life Science||b.Henan Animal and Poultry Health Products Engineering Technology Research Center,Nanyang Normal University,Nanyang Henan 473061,China||2.Department of Modern Service Management,Nanyang Vocational College of Science and Technology,Nanyang Henan 474150,China@@@1.a.School of Life Science||b.Henan Animal and Poultry Health Products Engineering Technology Research Center,Nanyang Normal University,Nanyang Henan 473061,China||2.Department of Modern Service Management,Nanyang Vocational College of Science and Technology,Nanyang Henan 474150,China
生物科学
猪圆环病毒4型Cap蛋白原核表达多克隆抗体
porcine circovirus type 4Cap proteinprokaryotic expressionpolyclonal antibody
《南阳师范学院学报》 2026 (1)
53-59,7
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