Cloning and Sequence Analysis of Interleukin 10(IL-10) Full-length cDNA from Cyprinus carpio L.

[Objective] This study aimed to obtain IL-10(interleukin 10) full-length cDNA of common carp(Cyprinus carpio L.) and conduct the sequence analysis.[Method] The differentially expressed cDNA fragment was obtained by DD-RTPCR(differential display RT-PCR) to get the full-length IL-10 cDNA.The cDNA library of peripheral blood leukocytes which were separated from common carp and stimulated with mitogen was screened by a probe labeled with DIG(digoxigenin).The IL-10 full-length cDNA was cloned from 0.8 ×104 recombinant phages,and the sequence analysis and homology comparison were carried out.[Result] Sequence analysis indicated that the IL-10 full-length cDNA of common carp was 1 117 bp long,containing a 55 bp 5′-UTR,a 522 bp 3′-UTR,and a 540 bp open reading frame(ORF) encoding 179 amino acids.In addition,there were three motifs for mRNA instability(ATTTA) in the 3'-untranslated region.The deduced protein sequence shared typical sequence features of the IL-10 family.Homology comparison indicated that the obtained sequence shared 89.1% homology with the carp IL-10 gene from GenBank.[Conclusion] This study laid foundation for further study of the expression manner,functional characteristic and regulation mechanism of IL-10 in vivo and the interaction mechanism in the inflammatory reaction and immune response.