红嘴鸥TLR7基因克隆与生物信息学分析

doi:10.16431/j.cnki.1671-7236.2022.01.005

Abstract

Abstract: [Objective] The Toll-like receptor 7 (TLR7) gene of Larus ridibundus was cloned, and its coding protein was analyzed by bioinformatics, so as to prepare for the study of antiviral activity of TLR7 protein in the later stage. [Method] The whole TLR7 gene sequence of Larus ridibundus was amplified by rapid amplification of cDNA ends (RACE). The TLR7 gene sequence, rare codon, similarity and physical and chemical properties, transmembrane region, signal peptide, N-glycosylation site, phosphorylation site and subcellular localization of TLR7 protein were analyzed by bioinformatics software. The secondary structure and tertiary structure of TLR7 protein were predicted by SOPMA and SWISS-MODEL softwares, respectively. [Result] The TLR7 gene of Larus ridibundus (GenBank accession No.:MZ668652) was successfully cloned. The total length of the gene sequence was 1 720 bp, the size of open reading frame (ORF) was 1 182 bp, and there were 39 rare codons, including 8 consecutive rare codons, encoding 393 amino acids. The similarity of TLR7 gene between Larus ridibundus and Gallus gallus, Tragopan temminckii, Coturnix coturnix, Anas platyrhynchos, Cygnus atratus, Paridae, Egretta garzetta, Aptenodytes forsteri, Gavia stellata, Pygoscelis papua in GenBank was 85.7%, 84.9%, 85.4%, 87.0%, 87.8%, 86.8%, 91.0%, 93.1%, 93.1% and 93.1%, respectively. The phylogenetic tree showed that it had the closest genetic relationship with Egretta garzetta. The molecular formula was C₂₀₈₀H₃₂₅₆N₅₅₆O₅₆₇S₁₉, the molecular weight of TLR7 protein was about 63 ku. The theoretical isoeletric point was 9.25. TLR7 protein had no transmembrane structure and signal peptide. It contained 6 N-glycosylation sites and 33 phosphorylation sites. It was a hydrophobic protein and mainly exists in the cytoplasm. The secondary structure of TLR7 protein was mainly alpha helix dominated, accounting for about 48.09%, followed by random coil (32.06%), extended chain (13.23%) and beta turn(6.62%). The tertiary structure of TLR7 protein was consistent with the secondary structure, and the similarity with the model protein human TLR7 protein was 68.78%. [Conclusion] The TLR7 gene of Larus ridibundus was closely related to the evolution of Egretta garzetta, it contained 8 consecutive rare codons and was difficult to express in vitro. The study provided an important reference for further exploring the antiviral immune mechanism of TLR7 protein in Larus ridibundus.

Key words: Larus ridibundus; Toll-like receptor 7 (TLR7); cloning; bioinformatics analysis

CLC Number:

S858.99

CHANG Hua, DUAN Gang, RUAN Qian, LUO Qianmin, YU Wanling, LIU Qingqi, HUANG Cuiqin, XIANG Xun. Cloning and Bioinformatics Analysis of TLR7 Gene in Larus ridibundus[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(1): 43-52.

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Cloning and Bioinformatics Analysis of TLR7 Gene in Larus ridibundus

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