Isolation and Analysis of Differential Expression Genes on Common Carp Peripheral Leucocyte Stimulated and Non-Stimulated with Mitogen by DDRT-PCR

DDRT-PCR is an effective and quick method to study differentially expressed genes on the same cells under differently physiological status and different stages. In this study fluorescence DDRT-PCR was used to compare mRNA of peripheral leucocytes of Common Carp stimulated and non-stimulated by mitogens (LPA, ConA and PHA) in different times such as 4 h, 12 h and 24 h. According to bioinformatic technology it indicated that it had 92 different fragments and 87 of them were re-amplified. After the gene was cloned and sequenced, 60 fragments were identified. The analysis of sequence and bioinformatic information revealed that the products of 5 cDNA fragments, which were highly homologous with thymosin beta , proteasome activator complex PA28 beta subunit, CXC chemokine, translation elongation factor-lbeta(EF-lbeta) and matrix metalloproteinase (MMPs) had immune response. The full-length thymosin beta cDNA was 528 bp with the open reading frame encoding 46 amino acid residues, and the sequences was submitted to GenBank to gain a accession number of AY457946.