China Animal Husbandry and Veterinary Medicine ›› 2022, Vol. 49 ›› Issue (7): 2474-2483.doi: 10.16431/j.cnki.1671-7236.2022.07.005

• Biotechnology • Previous Articles     Next Articles

Cloning, Bioinformatics and Tissue Expression Analysis of TRIM3 Gene in Swine

ZHANG Hang1,3, SHA Huiyang1,3, LI Huawei2, HUANG Liangzong1,3, ZHAO Mengmeng1,3   

  1. 1. College of Life Science and Engineering, Foshan University, Foshan 528231, China;
    2. College of Food and Bioengineering, Henan University of Animal Husbandry and Economics, Zhengzhou 450046, China;
    3. Veterinary Teaching Hospital, Foshan University, Foshan 528231, China
  • Received:2021-12-23 Online:2022-07-05 Published:2022-06-29

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Abstract: 【Objective】 Tripartite motif-containing 3 (TRIM3) gene of Large White pigs was cloned and analyzed by bioinformatics and tissue expression.【Method】 The full-length CDS sequence of TRIM3 gene in Large White pigs was amplified by PCR technology and cloned,pMD18-T vector was ligated and Escherichia coli DH5α competent cells were transformed,positive clones were screened by blue and white spots,sequenced after PCR identification of bacterial fluid,compared with TRIM3 gene sequences of different species and constructed phylogenetic tree,and bioinformatics analysis of its coding proteins was applied to a variety of online software.The relative expression of TRIM3 gene in different tissues of Large White pigs was detected by Real-time quantitative PCR.【Result】 The CDS sequence of TRIM3 gene in Large White pigs was 2 235 bp in total length and encoded 744 amino acids.Similarity and genetic evolution analysis results showed that the similarity between Large White pigs and Sus scrofa was the highest,up to 99.7%,and the similarity with Anas platyrhynchos was the lowest (75.1%).And the TRIM3 gene of Large White pigs was first clustered with Sus scrofa,and the relationship with Bos indicus and Capra hircus were closer.Bioinformatics analysis showed that the molecular mass of TRIM3 protein in Large White pigs was 80.58 ku,the theoretical isoelectric point (pI) was 8.32,and the instability coefficient was 40.85,which was a hydrophilic protein but not a secretory protein,and there were no glycosylation sites,which were predicted to have 60 phosphorylation sites,mainly in the cytoplasm;In the secondary structure of TRIM3 protein,random coil was the mainstay,accounting for 41.67%,and the prediction results of the tertiary structure model were consistent with the secondary structure.Tissue expression analysis showed that the TRIM3 gene in Large White pigs was distributed in heart,liver,spleen,lung,kidney,muscle,trachea and colon,and the expression in lung was the highest, and significantly higher than that in other tissues (P＜0.05).【Conclusion】 In this study,the full-length CDS sequence of TRIM3 gene in Large White pigs was successfully cloned and analyzed by bioinformatics and tissue expression,which provided a theoretical basis for the further study of the immunological function of TRIM3 protein in Large White pigs,and was of great significance to explore the molecular mechanism of TRIM3 gene in innate immunity and antiviral infection.

Key words: Large White pigs; TRIM3 gene; cloning; expression analysis

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