柔嫩艾美耳球虫MIF基因克隆及生物信息学分析

doi:10.16431/j.cnki.1671-7236.2023.06.029

Abstract

Abstract: 【Objective】 The purpose of this study was to obtain the CDS region of macrophage migration inhibitory factor (MIF) gene of Eimeria tenella, and explore its bioinformatics characteristics, so as to provide a theoretical basis for subsequent epitope screening.【Method】 The positive samples suspected of Eimeria tenella infection in Jinzhong area were collected for PCR identification.According to the MIF gene sequence of Eimeria tenella published in GenBank, the CDS region of MIF gene was amplified by RT-PCR, and the recombinant plasmid pET28a-EtMIF was constructed.PCR and double enzyme digestion were used for identification, sequencing and sequence analysis.Bioinformatics software was used to predict the physical and chemical properties, structure and function of MIF protein.【Result】 The CDS region of MIF gene in Eimeria tenella was successfully cloned, with a total length of 348 bp, encoding 115 amino acids, and a relative molecular weight was 1.203×10⁴.The phylogenetic tree analysis showed that the amino acid sequences of MIF gene of Eimeria tenella and Eimeria necatrix had the closest relationship, with the similarity of 98.3%.MIF protein was a non-transmembrane, soluble, non-secretory protein with subcellular localization mainly in the cytoplasm and contains 10 phosphorylation sites and a conserved domain.The secondary structure was composed of random coil (35.65%), extended chain (30.43%), alpha helix (27.83%) and beta turn (6.09%).The tertiary structure was predicted to be consistent with the secondary structure.There were three best potential epitopes.【Conclusion】 MIF gene of Eimeria tenella was successfully cloned, and the bioinformatics analysis of this gene was carried out.The results provided a reference for future research on the function of MIF gene and the screening of genetic engineering vaccine candidates.

Key words: Eimeria tenella; MIF gene; bioinformatics

CLC Number:

S852.72

LI Yongbin, YANG Huilin, SONG Zihao, ZHANG Yu, YANG Jin, MENG Jia, CUI Xiaozhen, LYU Xiaoling, ZHENG Mingxue, BAI Rui. Cloning and Bioinformatics Analysis of MIF Gene in Eimeria tenella[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2450-2459.

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Cloning and Bioinformatics Analysis of MIF Gene in Eimeria tenella

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