猪瘟病毒E2基因在Pichia pastoris中的表达及其免疫活性的初步研究

Study on the Expression of E2 Gene of Classical Swine Fever Virus in Pichia pastoris and the Immunological Activity of Its Expression Product

将猪瘟病毒的E2基因克隆入酵母分泌型表达载体pPIC9K中 ,酶切线性化后电穿孔导入Pichiapastoris进行整合 ,经G418筛选得到高拷贝转化子 ,甲醇诱导表达。SDS PAGE和Westernblot结果证实了酵母培养上清液中含有E2蛋白。免疫活性研究证明P .

E2 gene of classical swine fever virus (CSFV) was cloned into secretory pPIC9K Pichia pastoris expression vector. After being linearized by digestion , the vector was transformed into Pichia pastoris by electroporation to integrate with the genome, the transformants with high copies were screened by G418 and were induced to express with methonal. The results of SDS PAGE and Western blot demonstrated that the supernatant of the induced P.pastoris culture contained protein E2. The results of the study on the immunological activity indicated that the protein E2 expressed in P.pastoris can elicit animal bodies to produce antibodies against protein E2.