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浙江大学学报（农业与生命科学版）
浙江大学学报（农业与生命科学版）  2022, Vol. 48 Issue (2): 261-268    DOI: 10.3785/j.issn.1008-9209.2021.04.081
动物科学与动物医学     
猪的肝富集基因1a蛋白的分子鉴定(英文)
孙铭浩，黄妤琪，党燕娜，贺津
浙江大学动物科学学院，杭州 310058
Molecular characterization of pig LEG1a protein
Minghao SUN(),Yuqi HUANG,Yanna DANG,Jin HE()
College of Animal Sciences, Zhejiang University, Hangzhou 310058, China
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摘要：

新发现的肝富集基因1（live enriched-gene 1, LEG1）在斑马鱼肝脏发育及鸭嘴兽的先天性免疫应答过程中发挥着重要的调控作用，但是LEG1在真兽亚纲物种（如小鼠和人）中的功能鲜有报道。本研究以猪为模型，对猪的肝富集基因1a蛋白（pLEG1a）进行了分子鉴定。实验首先制备并验证了pLEG1a蛋白的兔源多克隆抗体，并验证了pLEG1a在猪唾液腺和肺部的表达。同时，在唾液中检测到了pLEG1a蛋白的信号，表明pLEG1a蛋白可能是一种分泌蛋白。此外，当在HEK293T细胞中异位表达pLEG1a时，可在细胞培养液中检测到pLEG1a蛋白，进一步证明了pLEG1a是一种分泌型蛋白。另外，糖基化实验显示，唾液及HEK293T细胞中的pLEG1a存在糖基化条带，且这一条带可以通过糖基化酶去除，表明pLEG1a是一种N-糖基化蛋白。综上所述，本研究探究了pLEG1a蛋白与其他物种（尤其是真兽亚纲动物）LEG1蛋白在表达谱、糖基化和分泌能力方面的相似性，并且提出pLEG1a可能是研究真兽亚纲物种LEG1基因的良好模型。
关键词： 猪;  肝富集基因1a蛋白;  分泌蛋白;  糖基化    
Abstract:

Liver-enriched gene 1 (LEG1) is a newly identified gene that plays an important role in the liver development of zebrafish and the innate immunity of platypus. However, little is known about LEG1 in eutherians such as mice and humans. In this study, we explored the molecular characteristics of the LEG1 protein in a pig model (pLEG1a). A rabbit polyclonal antibody against pLEG1a was produced and validated. Then, we detected the pLEG1a protein in both the salivary gland and lung. Moreover, pLEG1a could be detected in the saliva, suggesting its secretory nature. Ectopic expression of pLEG1a in HEK293T cells further confirmed that pLEG1a was a secretory protein. In addition, glycosylation tests showed the existence of glycosylated bands which could be enzymatically removed, indicating that the secreted pLEG1a was N-glycosylated. In conclusion, the pLEG1a protein is similar to the LEG1 proteins of other species, especially eutherian LEG1s, in terms of expression profile, glycosylation, and secretion ability, suggesting that pLEG1a is a good model to further study the function of eutherian LEG1s.
Key words: pig    LEG1a protein    secretory protein    glycosylation
出版日期: 2022-04-25
CLC:  S 828  
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孙铭浩
黄妤琪
党燕娜
贺津

引用本文:

孙铭浩, 黄妤琪, 党燕娜, 贺津. 猪的肝富集基因1a蛋白的分子鉴定(英文)[J]. 浙江大学学报（农业与生命科学版）, 2022, 48(2): 261-268.

Minghao SUN, Yuqi HUANG, Yanna DANG, Jin HE. Molecular characterization of pig LEG1a protein. Journal of Zhejiang University (Agriculture and Life Sciences), 2022, 48(2): 261-268.

链接本文:

https://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2021.04.081    或    https://www.zjujournals.com/agr/CN/Y2022/V48/I2/261

  
  
  
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