[High-level expression of the potential vaccine antigen TSO18 of Taenia solium in Pichia pastoris].
TSO18 gene was subcloned into the Pichia pastoris expression vector pPIC9K. The recombinant plasmid pPIC9K-TSO18 was transformed into P. pastoris GS115 by electroporation so that the plasmid will be integrated with chromosome of P. pastoris. The P. pastoris strains containing multi-copy recombinant were screened by G418 and induced by methanol. The expression product was analyzed by SDS-PAGE, Western blot, deglycosylation, and purified by Sephadex column, and was used to immunize mice. The results indicated that the target protein was efficiently expressed in P. pastoris, and glycosylated moderately, and had immunological activity. In a 5 liter fermentor, the expression level of the target protein was up to 2.54 mg/mL. These results will benefit for the development of genetically engineering vaccine.
Yuan
Gai-Ling
GL
Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China.
Cai
Xue-Peng
XP
Jing
Zhi-Zhong
ZZ
Zheng
Ya-Dong
YD
Luo
Xue-Nong
XN
Jia
Wan-Zhong
WZ
Li
Hui
H
Ding
Jun-Tao
JT
D000818:Animals; D000947:Antigens, Helminth / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D003001:Cloning, Molecular; D018274:Electroporation; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D051379:Mice; D010843:Pichia / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D013552:Swine; D041201:Taenia solium / Q000235:genetics* / Q000276:immunology*
D000947:Antigens, Helminth; D011994:Recombinant Proteins