Cloning and Sequence Analysis of the Main Cytokines in Bama Miniature Pig

In order to enrich the cytokine biological data of Guangxi Bama miniature pig,the lymphocytes of Bama miniature pig was separated from peripheral blood and the total RNA was extracted from the peripheral blood lymphocytes,respectively.The target genes of cytokine were amplified by RT-PCR and cloned into pMD18-T vector and transformed into E.coli DH5α,respectively.We chose positive clones and submitted them to the NCBI after sequencing the recombinant strain.Comparing homology of the cloned sequence with the known sequence,and using the software to analysis the cloned sequence.The results showed that the gene length of cloned cytokines IL-2,IL-12,IFN-γ,TNF-α of Bama miniature pig were 338,377,380,402 bp,respectively.It was consistent with the expected length of target gene.The sequences analysis showed that the homology of the cloned genes was relatively high with known domestic pigs or wild boars.The sequence homology of IL-2,IL-12,IFN-γ,TNF-α were 98.9%,99.5%,99.2%,100%,respectively.The research on cytokine gene sequence of Bama miniature pigs not only enriched the biological data of Bama miniature pigs,but also laid the foundation for research of the cytokine function.It also provided the basis for the quantitative detection of cytokine.