版纳微型猪近交系免疫排斥相关基因<i>B4GALNT2</i>的克隆、表达及功能生物信息学分析

doi:10.7668/hbnxb.20190232

摘要/Abstract

摘要： 为探讨B4GALNT2基因在异种器官移植排斥反应中的作用，以版纳微型猪近交系（Banna mini-pig inbred line，BMI）为试验动物，研究BMI B4GALNT2基因的序列、结构和表达等特性。通过RT-PCR方法从扁桃体组织中获得B4GALNT2基因cDNA序列，先后应用qPCR和Western Blotting分别检测其mRNA及蛋白质在多种组织中的表达情况，并对蛋白质结构特征进行相关生物信息学分析。获得BMI B4GALNT2 CDS序列1 509 bp，编码502个氨基酸（GenBank核酸登录号：KU358546；蛋白质登录号：ANH21179.1），功能生物信息学分析显示该蛋白含有半乳糖转移酶结构域，有一个跨膜结构，无信号肽。荧光定量结果显示其mRNA在扁桃体、脾脏和淋巴结等重要免疫组织中相对高表达，Western Blotting结果同样显示BMI B4GALNT2蛋白在扁桃体等免疫组织中相对高表达。明确了B4GALNT2基因在BMI多组织中的表达情况，以及B4GALNT2蛋白质的基本结构和功能特征，为进一步深入探究B4GALNT2基因在猪-人异种移植免疫排斥方面的分子机制奠定基础。

关键词: 版纳微型猪近交系, β1,4-N-乙酰氨基半乳糖转移酶2(B4GALNT2), 免疫排斥, 异种移植, 表达分析, 生物信息学分析

Abstract: In this study, the Banna mini-pig inbred line(BMI) was used as an experimental animal to study the sequence, structure and expression of BMI B4GALNT2 gene, and to explore the role of B4GALNT2 gene in xenotransplant rejection. The cDNA sequence of B4GALNT2 gene was obtained from tonsil tissue of Banna mini-pig inbred line(BMI) by RT-PCR and the expression levels of mRNA and protein in various tissues were detected by qPCR and Western Blotting, respectively. Further, the structure features of B4GALNT2 protein were analyzed using bioinformatics. A CDS sequence of 1 509 bp of BMI B4GALNT2 was obtained, with a GenBank nucleotide accession number KU358546 and a protein accession number ANH21179.1, which encoded 502 amino acids. Functional bioinformatics analysis revealed that B4GALNT2 protein contained one galactose transferase domain and one transmembrane structure without signal peptide. Fluorescence quantitative results indicated that B4GALNT2 mRNA expressed highly in some important immune tissues, such as tonsils, spleen and lymph nodes. Meanwhile, Western Blotting results also showed that BMI B4GALNT2 protein was relatively highly expressed in tonsil and other immune tissues.The expression of B4GALNT2 gene in BMI multitissue and the basic structure and functional characteristics of B4GALNT2 protein were clarified, laying a foundation for further molecular mechanism study of the B4GALNT2 gene on the immune rejection in the process of xenotransplantation from pig to human.

Key words: Banna mini-pig inbred line, β1,4-N-acetylgalactosyltransferase2(B4GALNT2), Immune rejection, Xenotransplantation, Expression analysis, Bioinformatics analysis

中图分类号:

Q78
S828.2

宋雪, 王淑燕, 陈园园, 王配, 霍海龙, 张霞, 霍金龙. 版纳微型猪近交系免疫排斥相关基因B4GALNT2的克隆、表达及功能生物信息学分析[J]. 华北农学报, 2019, 34(6): 219-226. doi: 10.7668/hbnxb.20190232.

SONG Xue, WANG Shuyan, CHEN Yuanyuan, WANG Pei, HUO Hailong, ZHANG Xia, HUO Jinlong. Cloning, Expression and Functional Bioinformatics Analysis of B4GALNT2 Gene Related to Immune Rejection in Banna Mini-pig Inbred Line[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2019, 34(6): 219-226. doi: 10.7668/hbnxb.20190232.

http://www.hbnxb.net/CN/Y2019/V34/I6/219

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版纳微型猪近交系免疫排斥相关基因B4GALNT2的克隆、表达及功能生物信息学分析

Cloning, Expression and Functional Bioinformatics Analysis of B4GALNT2 Gene Related to Immune Rejection in Banna Mini-pig Inbred Line

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