现代食品科技2018，Vol.34Issue(4)：51-55,5.DOI:10.13982/j.mfst.1673-9078.2018.04.009
重组猪圆环病毒2型Cap蛋白在毕赤酵母中的优化表达及疫苗活性研究
Optimized Expression and Vaccine Activity of Recombinant Porcine Circovirus Type 2 Capsid Protein in Pichia Pastoris
摘要
Abstract
Porcine circovirus type2 (PCV2) is the major pathogen of weaning multi-systemic wasting syndrome (PMWS), which has caused significant economic losses to the swine industry worldwide. Porcine circovirus type 2 capsid protein (PCV2-Cap) is the main target protein for the preparation of vaccine against weaning multi-systemic wasting syndrome (PMWS). In this study, the recombinant expression and vaccine activity of PCV2-Cap protein in Pichia pastoris were investigated to solve the problems of complex production process and high cost of PCV2 vaccine. After removing the nuclear localization sequence (NLS) from PCV2-Cap gene sequence, a preferential optimization of codon was performed to construct a recombinant expression vector and transformed into the host bacteria (X-33) of P. pastoris. The results of SDS-PAGE and Western blot showed that the Cap protein was successfully expressed in the yeast expression system with a protein size of 26 Ku, and the high density fermentation test in 5 L bioreactor suggested that the concentration of PCV2-Cap protein in fermentation supernatant was 0. 53 mg/m L, accounting for 23. 5％ of the total cell protein. In addition, the animal immunization test demonstrated that Cap protein could stimulate the body to produce specific antibodies. Consequently, the PCV2-Cap gene was secreted and expressed P. pastoris, which might make a contribution to the development of PCV2-Cap subunit vaccines.关键词
猪圆环病毒2型/毕赤酵母/重组表达/疫苗活性Key words
porcine circovirus type 2/Pichia pastoris/recombinant expression/vaccine activity引用本文复制引用
王斌,罗时渝,谢景毅,叶燕锐,潘力..重组猪圆环病毒2型Cap蛋白在毕赤酵母中的优化表达及疫苗活性研究[J].现代食品科技,2018,34(4):51-55,5.基金项目
广东省科技计划项目(2016A050503016、2016A010105004) （2016A050503016、2016A010105004）
广州市科技计划项目(201510010191) （201510010191）
广东省自然科学基金项目(2017A030313097) （2017A030313097）
