[Expression and characterization of mouse 17-1A antigen from Pichia pastoris].
OBJECTIVE
This is a ground work in preparation for the development of a vaccine against tumor, in which 17-1A glucoprotein, a tumor associated antigen (TAA), was cloned and expressed in Pichia pastoris.


METHODS
The cDNA of 17-1A gene was cloned from nephridial tissue of mouse embryo by RT-PCR and subsequently inserted into pPICZaA plasmid after digestion with Xho I and Xba I. The linearized recombinant plasmid was transformed into Pichia pastoris strain GS115. 17-1A glucoproteins were secreted into culture supernatant through alpha-signal peptide of AOX I after inducation with methanol. The protein products were analyzed and characterized by SDS-PAGE and Western-blot.


RESULTS
The plasmid pPICZalphaA-17-1A was constructed and verified by sequenceing. 17-1A glucoproteins were expressed stably and glycosylated in Pichia pastoris.


CONCLUSIONS
Glycosylated tumor associated antigen 17-1A can be expressed in Pichia pastoris.
Wang
Yong-sheng
YS
State Key Laboratory of Biotherapy, Cancer Center, West China Hospital, Sichuan University, Chengdu 610041, China.
Tian
Ling
L
Diao
Peng
P
Hu
Min
M
Du
Xiao-bo
XB
Wei
Yu-quan
YQ
D000818:Animals; D000951:Antigens, Neoplasm / Q000096:biosynthesis* / Q000235:genetics; D019496:Cancer Vaccines / Q000276:immunology; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004622:Embryo, Mammalian; D007668:Kidney / Q000737:chemistry; D051379:Mice; D010843:Pichia / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics*
D000951:Antigens, Neoplasm; D019496:Cancer Vaccines; D018076:DNA, Complementary; C050928:carcinoma-associated antigen 17-1A