Identification of immune responsible fibrinogen beta chain in the liver of large yellow croaker using a modified annealing control primer system.
In this article, we used a modified ACP system (mACP) developed in our laboratory to analyze differentially expressed genes in the liver of large yellow croaker, Pseudosciaena crocea (Richardson). By using 20 pairs of mACPs, 7 differentially expressed genes were obtained. One of the genes we identified encodes for a fibrinogen beta chain (FGB). The full-length cDNA of FGB was 1645 bp, including 5 bp of 5' untranslated region (5'-UTR), 1479 bp of open reading frame (ORF), and 161 bp of 3'-UTR. The ORF was capable of encoding 492 amino acids with an estimated molecular mass of 55.6 kDa, giving it a predicted pI of 5.94. The deduced amino acid sequence included an FGB profile (V(238)-Y(488)) and an FGB family signature (WWYNRCHSANPNG). Multiple sequence alignments indicated that the large yellow croaker FGB showed homology with FGB sequences of other species (45-77% identity). Real time PCR analysis demonstrated that the expression of FGB in the liver of large yellow croaker injected with Vibrio parahaemolyticus was significantly (P < 0.05) lower than that of the control group at 8 d, which confirmed the expression patterns of the results of mACP differential display.
Xie
F J
FJ
The Key Laboratory of Science and Technology for Aquaculture and Food Safety, Fisheries College, Jimei University, Xiamen, Fujian 361021, China.
Zhang
Z P
ZP
Lin
P
P
Wang
S H
SH
Zou
Z H
ZH
Wang
Y L
YL
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers; D005340:Fibrinogen / Q000737:chemistry / Q000235:genetics*; D005786:Gene Expression Regulation; D008099:Liver / Q000737:chemistry / Q000276:immunology*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization / Q000379:methods; D010473:Perciformes / Q000235:genetics* / Q000276:immunology*; D010802:Phylogeny; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D017931:DNA Primers; D005340:Fibrinogen