羊IL-1Ra基因全长cDNA克隆及生物信息学分析

doi:10.16431/j.cnki.1671-7236.2018.03.005

Abstract

Abstract:

This study was aimed to clone ovine interleukin-1 receptor antagonist (IL-1Ra) gene and analyze the gene structure using bioinformatics.The primers was designed according to the obtained SSH cDNA library IL-1Ra gene sequence information and the known nucleotide sequence(GenBank accession No.:KC425613.1),IL-1Ra gene was amplified and cloned by RT-PCR and RACE methods,IL-1Ra sequence and its related molecular characteristics were analyzed.The results showed that the length of IL-1Ra gene was 1 228 bp,and encoded 174 amino acids.It was predicted that IL-1Ra had a complete IL-1 conserved domain and IL-1Ra domain (PHA02651 domain).The molecular weight of IL-1Ra was 19 765.8 u,the isoelectric point (pI) was 5.72,its molecular was C₈₈₅H₁₃₈₅N₂₃₅O₂₅₆S₁₁.IL-1Ra contained the signal peptide.The secondary structure of IL-1Ra had more β-fold and receptor binding sites,which were consistent with the predicted results of tertiary structure,and had a similarity to the spatial structure of human/mouse IL-1Ra,up to more than 90%.The IL-1Ra contained a clover structure which was peculiar to the IL-1,and its enzyme-binding domain was located between TYR⁴⁷ and GLU⁶⁶.The results showed that there were five highly homologous cysteines between sheep and the other fourteen species.By establishing the phylogenetic tree of IL-1Ra,it was found that the full-length cDNA of IL-1Ra in sheep formed a branch with goat and cattle.This study result could provide the basis for the further study of the biological function of IL-1Ra gene.

Key words: sheep; IL-1Ra gene; bioinformatics; analysis

CLC Number:

Q785

SHUI Yiming, LIU Nannan, GUO Xing, ZOU Deying, ZHAI Feifei, WANG Lulu, JU Dandi, ZHANG Shijun, HU Pan, LI Yansong, LU Shiying, ZHOU Yu, LIU Zengshan, LI Zhaohui, REN Honglin. Cloning and Bioinformatics Analysis of IL-1Ra Gene Full-length cDNA in Sheep[J]. , 2018, 45(3): 598-603.

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Cloning and Bioinformatics Analysis of IL-1Ra Gene Full-length cDNA in Sheep

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