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

摘要

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 4h, 12h and 24h. 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 β, proteasome activator complex PA28 β subunit, CXC chemokine, translation elongation factor-1β(EF-1β) and matrix metalloproteinase (MMPs) had immune response. The full-length thymosin β 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.