Screen And Study Of Novel Antimicrobial Peptide Genes From Penzer Crucian

Antimicrobial peptides (AMPs) are a kind of small size molecular peptides encoded by a special gene, with complex structures and components. AMPs are a very important part of organism's natural immunity. With unique antimicrobial mechanism, a broad spectrum of bactericidal effect, high antibacterial efficacy and hard to be resist by target microbial, it causes people's extensive concern. The development and application of AMPs is the focus of research and exploitation, in the pass few years. A large number of AMPs have been found in many kinds of organism, like: insects, mammals, amphibian, but few from fresh water fish. The aim of this research is that isolation and screening of new AMPs gene from panzer crucian, and analysis the AMPs' structure, function and mechanism of action. Prepare for designing and expression new AMPsTo get the gene of AMPs, we induced panzer crucian by Aeromonas hydrophila, and extract RNA from the liver. The different fragments were found in DDRT-PCR actions, fragments that length around 150bp were extracted, amplified and then sequenced. According to the new sequences, we designed primers and use RACE action to find the full sequence of the different fragments.We used 2 anchor primers matched pairs with 8 random primers in the DDRT-PCR action, and found many different fragments. The fragments that fit in length were sequenced,and got 4 sequences : CF-1,CF-2,CF-3,CF-4. Sequence Alignment of this sequences in the Genbank,we found that:CF-1,CF-2 is very conservative in the progress of evolution,they are windly present in many kinds of species.And CF-4 homologous sequence exists in the Otolemur garnetti genome.The function of this three sequence has not been reported at present, they maybe the novel antimicrobial peptide genes from penzer crucian.