Serum amyloid A in geese; cloning and expression of recombinant protein.
We defined the nucleotide-sequence of the full-length goose serum amyloid A and compared it to SAA sequences of the duck. The aim of this work was to clone and express recombinant goose SAA and to produce antibody against this protein: Total RNA was isolated from goose liver and used to synthesise first strand cDNA. The coding region of the goose SAA cDNA was amplified by PCR using primers corresponding to the appropriate conservative regions of duck SAA mRNA. The product was subcloned into pET-15b expression vector to result in a His*Tag fusion protein expression. The protein was purified by affinity chromatography. Rabbits were then immunized against the recombinant purified goose SAA protein. The anti-SAA serum was tested by Western blotting. Full-length goose SAA mRNA sequence has been obtained and sequenced.
Kovács
Beáta Marianna
BM
Department of Physiology and Biochemistry, Faculty of Veterinary Science, Szent István University, Budapest, Hungary.
Szilágyi
László
L
Janan
Janbaz
J
Rudas
Péter
P
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004591:Electrophoresis, Polyacrylamide Gel; D005777:Geese; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D000685:Serum Amyloid A Protein / Q000737:chemistry / Q000235:genetics*
D018076:DNA, Complementary; D012333:RNA, Messenger; D011994:Recombinant Proteins; D000685:Serum Amyloid A Protein