Cloning and sequence analysis of IL-4 gene in giant panda

By reverse transcription polymerrase chain reaction(RT-PCR),giant panda IL-4 gene was cloned from total RNA which was extracted from ConA-stimulated-cultured peripheral blood lymphocytes of giant panda,the amplified products of PCR were cloned into PGEM-T vector.After identification and sequencing,the sequence analysis revealed that the open reading frame of giant panda IL-4 gene contained 396 nucleotides which encoding 132 amino acids,the signal peptide contained 24 amino acids and the mature IL-4 gene was composed of 108 amino acids.Compared with one published sequence of giant panda(DQ166512),the homology of nucleotide sequence and deduced amino acid sequence were 99.5%and 100% respectively;compared with other mammals in GenBank,including dog,cat,human,pig,cow,horse,goat,rabbit,rat and so on,the homology of nucleotide sequence was between 50.7%(rat)-87.7%(dog),the homology of amino acid sequence was between 35.7%(rat)-78.8%(dog).Phylogenetic analysis based on the nucleotide sequence of the IL-4 gene showed that giant panda had the most closed genetic distance with dog and cat.