[Construction and expression of the Ts87 gene of Trichinella spiralis in Pichia pastoris].
OBJECTIVE
To construct the recombinant plasmid P(PICZ) alphaA-Ts87 and obtain secretive protein in Pichia pastoris.


METHODS
Ts87 was amplified by PCR and cloned into the expression vector P(PICZ) alphaA of P. pastoris to form P(PICZ) alphaA-Ts87. After the recombinant was isolated and linearized, it was transformed into P. pastoris GS115 by EasyComp kit and screened for zeocin resistant with different conditions and Mut phenotype. The high resistant Mut+ clones were cloned and the product induced by methanol was tested by dot hybridization. The expression product was further identified by SDS-PAGE and Western blotting.


CONCLUSIONS
The recombinant plasmid P(PiCZ) alphaA-Ts87 has been constructed. Ts87 gene has been expressed and secreted in Pichia pastoris GS115 strain.
Wang
Shao-hua
SH
Department of Parasitology, School of Basic Medicine, Capital University of Medical Sciences, Beijing 100054, China.
Zhu
Xin-ping
XP
Gu
Yuan
Y
Yang
Ya-ping
YP
Yang
Jing
J
D000818:Animals; D000947:Antigens, Helminth / Q000235:genetics* / Q000378:metabolism; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D010843:Pichia / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014170:Transformation, Genetic; D017160:Trichinella spiralis / Q000235:genetics*
D000947:Antigens, Helminth; D011994:Recombinant Proteins