猪Btnl5基因的克隆、表达分析及对NF-κB信号通路的调控作用

doi:10.13560/j.cnki.biotech.bull.1985.2016-1151

生物技术通报 ›› 2017, Vol. 33 ›› Issue (6): 142-148.doi: 10.13560/j.cnki.biotech.bull.1985.2016-1151

猪Btnl5基因的克隆、表达分析及对NF-κB信号通路的调控作用

包琦^1,2,3, 赵凯¹, 段子渊²

1. 中国科学院西北高原生物研究所高原生物适应与进化重点实验室中国科学院西北高原生物研究所高原鱼类进化与功能基因组学实验室,西宁 810001;
2. 中国科学院遗传与发育生物学研究所遗传资源中心,北京 100101;
3. 中国科学院大学,北京 100049

收稿日期:2016-12-20 出版日期:2017-06-26 发布日期:2017-06-19
作者简介:包琦,女,博士研究生,研究方向：分子生物学;E-mail：baoqi1988@126.com
基金资助:
国家重点基础研究发展计划（“973”计划）（2013CB835200）

Cloning,Expression Analysis and Regulation to NF-κB Pathway of Btnl5 in Porcine

BAO Qi^1,2,3, ZHAO Kai¹, DUAN Zi-yuan²

1. Key Laboratory of Adaptation and Evolution of Plateau Biota,Northwest Institute of Plateau Biology,Chinese Academy of Sciences,Laboratory of Plateau Fish Evolutionary and Functional Genomics,Northwest Institute of Plateau Biology,Chinese Academy of Sciences,Xining 810001;
2. Genetic Resource Center,Institute of Genetics and Developmental Biology,Chinese Academy of Sciences,Beijing 100101;
3. University of Chinese Academy of Sciences,Beijing 100049

Received:2016-12-20 Published:2017-06-26 Online:2017-06-19

摘要/Abstract

摘要：

猪类嗜乳脂蛋白5（Btnl5）属于嗜乳脂蛋白家族,其同源蛋白多为免疫调节蛋白,为探索Btnl5的序列特征及其功能,利用反转录PCR（RT-PCR）和cDNA末端快速扩增（RACE）技术对Btnl5基因cDNA全长进行克隆及序列分析,采用定量PCR技术检测Btnl5在猪不同组织中的表达丰度,利用报告基因法检测Btnl5对NF-κB信号通路的影响。克隆得到Btnl5基因cDNA全长共3 334 bp,包含1 680 bp的开放读码框,可编码长度为559个氨基酸残基的多肽链。蛋白结构预测表明Btnl5编码的蛋白含有信号肽、两段免疫球蛋白结构域、跨膜结构域和B30.2结构域。定量PCR分析表明,在检测的11个组织中,Btnl5只在空肠中具有较高水平的表达。报告基因检测结果显示Btnl5对NF-κB信号通路有一定的抑制作用。研究结果说明,Btnl5可能通过抑制NF-κB信号通路参与调节肠道免疫。

关键词: 猪, 类嗜乳脂蛋白5, 表达分析, 报告基因检测, 核因子κB

Abstract:

Porcine butyrophilin-like 5（Btnl5）belongs to butyrophilin family and most of its homologous proteins are immune regulators. In order to explore the sequence characteristics and function of Btnl5,full-length cDNA of Btnl5 was cloned and characterized by reverse transcription PCR and rapid amplification of cDNA ends（RACE）. Then the mRNA expression abundances of Btnl5 in different tissues were detected using quantitative PCR,and its effect on NF-κB pathway was detected by reporter gene assay. The full length cDNA of Btnl5 was 3 334 bp containing a 1 680 bp open reading frame（ORF）encoding 559 amino acids. Structural analysis showed that this protein was a transmembrane protein,and its extracellular region contained a signal peptide,two immunoglobulin domains,its intracellular region was B30.2 domain. Quantitative PCR analysis showed that Btnl5 transcripts almost exclusively expressed in jejunum among 11 detected tissues. Reporter gene analysis indicated that Btnl5 suppressed the NF-κB pathway. The above results suggest that Btnl5 may involve in intestinal immune regulation through inhibiting NF-κB signaling pathway.

Key words: porcine, butyrophilin-like 5, expression analysis, reporter gene assay, nuclear factor κB

包琦, 赵凯, 段子渊. 猪Btnl5基因的克隆、表达分析及对NF-κB信号通路的调控作用[J]. 生物技术通报, 2017, 33(6): 142-148.

BAO Qi, ZHAO Kai, DUAN Zi-yuan. Cloning,Expression Analysis and Regulation to NF-κB Pathway of Btnl5 in Porcine[J]. Biotechnology Bulletin, 2017, 33(6): 142-148.

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猪Btnl5基因的克隆、表达分析及对NF-κB信号通路的调控作用

Cloning,Expression Analysis and Regulation to NF-κB Pathway of Btnl5 in Porcine

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