鸡FKBP5基因的克隆、生物信息学及组织表达谱分析

doi:10.16431/j.cnki.1671-7236.2021.01.004

Abstract

Abstract: The purpose of this study was to perform cloning and bioinformatics analysis of FK506 binding protein 5(FKBP5) gene in chickens,and detect its expression of different tissues in chickens.The spleen tissue cDNA in broiler was used as a template,the complete CDS region of FKBP5 gene in broilers was amplified and cloned,the homology was compared and phylogenetic tree was constructed.The physical and chemical properties,hydrophobicity,transmembrane structure,signal peptide,secondary structure and tertiary structure of FKBP5 protein were analyzed by online software.Real-time quantitative PCR was used to detect the expression of FKBP5 gene in different tissues of valgus-varus deformity(VVD) broilers and normal broilers.The results showed that the total CDS region sequence of FKBP5 gene in chickens was 1 350 bp,which encoded 449 amino acids.The homology of amino acid sequence of FKBP5 in chickens with Coturnix japonica,Anas platyrhynchos,Gekko japonicus,Pelodiscus sinensis,Mus musculus,Homo sapiens,Sus scrofa and Danio rerio were 98.4%,96.2%,85.8%,87.4%,81.1%,85.2%,86.1% and 61.2%,respectively.The genetic relationship with Coturnix japonica and Anas platyrhynchos was the closest,followed by reptiles and mammals,and it had the farthest relationship with Danio rerio.The molecular weight of chicken FKBP5 protein was 50.43 ku,its theoretical isoelectric point was 5.94,the half-life was 30 h,the N-terminus of the peptide chain was methionine,the instability coefficient was 23.80,belonging to the stable protein.The prediction results of transmembrane region and signal peptide showed that FKBP5 protein was no transmem brane structure and signal peptide.Domain analysis showed that the protein contained two FKBP-type peptidylprolyl isomerases and three tetrapeptide repeats (TPR).FKBP5 protein structure was mainly consisted of alpha helix (46.77%),extended chain (12.47%) and random coil (40.76%),it had strong correlation with HSPA2,PPID,STIP1,HSP90AA1 and HSP90AB1.Real-time quantitative PCR results revealed that FKBP5 gene in chickes was expressed in all tissues.The expression of tissues in VVD broilers were higher than that of normal broilers,and the expression in heart,leg muscle,bursa,thymus and cartilage was significantly different (P<0.05),the expression in spleen was extremely significantly different (P<0.01).The results of this experiment provided reference for broiler bone disease and leg healthy breeding.

Key words: chicken; FKBP5 gene; cloning; bioinformatics analysis; tissue expression analysis

CLC Number:

TANG Hehe, ZHANG Zhenzhen, ZHANG Xianzhen, JIANG Ruirui, LI Guoxi, TIAN Yadong, KANG Xiangtao, HAN Ruili. Cloning,Bioinformatics and Tissue Expression Profile Analysis of FKBP5 Gene in Chickens[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(1): 35-43.

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Cloning,Bioinformatics and Tissue Expression Profile Analysis of FKBP5 Gene in Chickens

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