[Expression and glycosylation of rotavirus strain SA11 VP4 protein in a recombinant adenovirus].
OBJECTIVE
A modified VP4 gene of rotavirus SA11 strain was expressed by a recombinant human adenovirus.


METHODS
A whole VP4 gene was obtained with PCR and induced the signal peptide at the gene N terminal. The chimera gene was cloned into pCMV plasmid that consist of human cytomagolovirus promoter and then cloned the gene to transfer human adenovirus type 5 vector. Homologues recombinant was performed by co-transfection to 293 cell line with recombinant plasmid and viral genome using CaPO4 precipitation.


RESULTS
VP4 gene is 2,362 base pair in length mutation was not found in whole VP4 gene sequence. Expressed product in recombinant adnovirus was confirmed to be specific and more antigenicity by indirect immunofluorescence assay. Both Western blot and immunoprecipitation assays showed that expressed protein molecular weight was higher than wild type VP4 protein and that modified product was corresponding to a glycosylation of VP4 protein.


CONCLUSIONS
It may be a effective method to modify interested gene for enhancing stability, antigenicity and immunogenicity of expressive product.
Sun
M
M
Department of Molecular Biology, Institute of Medical Biology, CAMS, PUMC, Kunming 650107, China.
Zhang
Y
Y
Ma
Y
Y
Zhang
G
G
Du
Q
Q
Dai
C
C
D000260:Adenoviruses, Human / Q000235:genetics*; D036022:Capsid Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D005822:Genetic Vectors; D006031:Glycosylation; D011994:Recombinant Proteins / Q000096:biosynthesis; D014162:Transfection
D036022:Capsid Proteins; D011994:Recombinant Proteins; C060339:VP4 protein, Rotavirus