The Cloning And Sequence Analysis Of The CDNA Of Camel β-Defensin CaBD-1

Antimicrobial peptides are important components of the innate immune system.The are microbicidal agents and can protect living body.Defensins constitute a large family of antimicrobial peptides whose members have isolated from species throughout the plant and animal kingdoms.They demonstrate effective and broad-spectrum antimicrobial activity.The have special antimicrobial mechanism different from antibiotic which can not induce germ variation.Based on molecular architecture and different source,defensins are divided asα-defensin ,β-defensin,θ-defensin,insect defensin and plant defensin. Here we report the identification of a novel β–defensin from camel tisses,camel β–defensin-1(caBD-1). Based on the conserved sequence of other cud chewer,the primer were designed. Total RNA was extracted from the tongue epithelia of camel and the cDNA encoding caml β–defensin (caBD-1) was amplified by the reverse transcription PCR(RT-PCR). The purified RT-PCR product was cloned in pBlueselct T vector. The results of restriction endonuclease pattern analysis of recombinant plasmid and DNA sequencing demonstrated that camelβ–defensin caBD-1 is belong to the family of β–defensin because the cDNA contain a Open Reading Frame(ORF) of 192 bases which encoded a 64 amino acid prepro-peptide and the prepro-peptide contained the β–defensin consensus sequence of six conserved cysteine residues. RACE technic is shortened form of the technic of Rapid Amplication of cDNA Ends.The anchored-PCR-RACE strategy was used to amplify 3ˊend of cDNA and backward nested- PCR-RACE strategy was used to amplify 5ˊend of cDNA.Then,they were cloned and were identified to encode the full cDNA sequence of caBD-1...