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.
The Key Laboratory of Science and Technology for Aquaculture and Food Safety, Fisheries College, Jimei University, Xiamen, Fujian 361021, China.
F J
FJ
Xie
Z P
ZP
Zhang
P
P
Lin
S H
SH
Wang
Z H
ZH
Zou
Y L
YL
Wang
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