CAJ | 학술논문

将禽白血病病毒p27基因克隆并构建重组表达质粒pET28a-p27，在大肠杆菌BL21中经IP TG诱导后产生可溶性表达蛋白。表达的蛋白用Western blot进行活性检测，其能与辣根过氧化物酶标记的兔抗p27抗体发生特异性反应；采用金属螯和层析对表达蛋白进行纯化，其纯度约为95％；用纯化的表达蛋白p27免疫家兔制备抗血清，ELISA抗体效价可达1∶25600。研究结果表明，大肠杆菌表达的p27蛋白具有良好的抗原性，可以替代纯化病毒蛋白用于禽白血病病毒检测。
장금백혈병병독p27기인극륭병구건중조표체질립pET28a-p27，재대장간균BL21중경IP TG유도후산생가용성표체단백。표체적단백용Western blot진행활성검측，기능여랄근과양화물매표기적토항p27항체발생특이성반응；채용금속오화층석대표체단백진행순화，기순도약위95％；용순화적표체단백p27면역가토제비항혈청，ELISA항체효개가체1∶25600。연구결과표명，대장간균표체적p27단백구유량호적항원성，가이체대순화병독단백용우금백혈병병독검측。
p27 protein gene from avian leukosis virus was cloned and constructed with plasmid pET-28a(+) . The recombinant protein p27 was expressed in an E. coli strain BL21(DE3) with IPTG induction and detected by Western blot with horseradish peroxidase labeled rabbit-anti-p27 antibodies. After that,the protein was purified to above 95% of purity by affinity chromatography with Ni-NTA agarose column. The anti-sera against p27 protein was obtained by immunizing rabbit with the purified recombinant p27 protein. The specificity of polyclonal antibody was about 1 ∶ 256000 after detected by ELISA. The results show that the p27 protein expressed by E. coli has good antigenicity and can replace the purified virus protein for detection of avian leukosis virus.