鸡DCAF1生物信息学分析及对J亚群禽白血病病毒复制的影响

doi:10.16431/j.cnki.1671-7236.2024.05.005

Abstract

Abstract: 【Objective】 The purpose of this study was to analyze the protein characteristics of chicken DDB1 and CUL4 associated factor 1(DCAF1) and construct an eukaryotic expression vector,with investigating the effect of DCAF1 on ALV-J replication.【Method】 The amino acid sequence of DCAF1 protein of related species were downloaded from NCBI and a phylogenetic tree was constructed using Mega 11.0 software.Online programs such as ProtParam,ProtScale,and TMHMM were utilized to analyze the amino acid sequence of DCAF1 protein in chicken,predict its biological information such as physicochemical properties,conserved domains,secondary and tertiary structures,and interacting proteins.The recombinant eukaryotic expression vector of DCAF1 gene in chicken was constructed,and the effects of ALV-J infection on the expression of DCAF1 mRNA and ALV-J Env protein,as well as the effects of overexpression of DCAF1 on ALV-J replication were detected using Real-time quantitative PCR and Western blotting techniques.【Result】 The DCAF1 protein of chicken was about 1 496 amino acids,and its molecular weight was about 170 ku. The results of phylogenetic tree showed the chicken was closet related to Oxyura jamaicensis and farthest related to Danio rerio.The bioinformatics prediction results indicated that DCAF1 protein of chicken had no transmembrane domain,no signal peptide expression,and contained three main structural domains:ARM folding,LisH and WD40 duplication,and one acidic domain at the end,and interacted with proteins such as DDB1,CUL4A and SAMHD1.The secondary structure of DCAF1 protein of chicken was mainly consisted of random coil and alpha helix,which accounted for 44.85% and 41.18%;followed respectively by extended chain and beta turn,which accounted for 9.76% and 4.21%.The tertiary structure results were basically consistent with the secondary structure,with a modeling consisitency of 93.79%.After 24 and 48 h of ALV-J infection,the protein and gene expression of DCAF1 were extremely significantly higher than those at 0 h of infection (P＜0.01).After overexpression of DCAF1,compared with the infection control group,the ALV-J Env protein was significantly increased (P＜0.05).【Conclusion】 The DCAF1 protein of chicken consisted of 1 496 amino acids with a molecular weight of about 170 ku.It was a stable hydrophilic protein and did not contain a transmembrane domain.This study successfully constructed eukaryotic expression vector of DCAF1 gene.In ALV-J infected cells,the expression of DCAF1 protein and gene were extremely significantly increased,while overexpression of DCAF1 significantly promoted ALV-J replication.This results laid the foundation for further elucidating the biological function of chicken DCAF1 and its molecular mechanism of interaction with ALV-J.

Key words: chicken; DCAF1; ALV-J; eukaryotic expression vector; bioinformatics

CLC Number:

S831

PAN Shiyu, YIN Na, WANG Jiaxing, WANG Qiangzhou, CHEN Shihao, BAI Hao, CHANG Guobin. Bioinformatic Analysis of Chicken DCAF1 and Its Impact on the Replication of ALV-J[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(5): 1837-1845.

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Bioinformatic Analysis of Chicken DCAF1 and Its Impact on the Replication of ALV-J

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