학술논문
太行鸡plekhm2基因结构与差异表达分析 / Analysis of plekhm2 Gene Structure and Differential Expression in Taihang Chickens(Gallus domesticus)
Document Type
Academic Journal
Author
Source
农业生物技术学报 / Journal of Agricultural Biotechnology. 32(1):180-188
Subject
Language
Chinese
ISSN
1674-7968
Abstract
鸡沙门氏菌病是由沙门氏菌(Salmonella)感染鸡(Gallus domesticus)所引起的一种传染性极高的细菌性疾病,含pleckstrin同源性和RUN结构域M2(pleckstrin homology and RUN domain containing M2,plekhm2)基因作为沙门氏菌效应蛋白SifA的靶点,通过下调驱动蛋白的募集在沙门氏菌侵染中发挥作用.本研究旨在分析鸡plekhm2基因的序列特征,并研究其在太行鸡中的组织表达谱以及在感染和未感染沙门氏菌太行鸡肝脏和脾脏中的表达水平.基于沙门氏菌感染和未感染鸡的脾脏转录组筛选差异表达基因,利用在线工具和软件对plekhm2基因进行生物信息学分析,利用qPCR技术检测plekhm2基因在太行鸡不同组织的表达水平,并构建太行鸡沙门氏菌感染模型研究plekhm2在肝脏和脾脏中的表达变化.结果表明,通过脾脏转录组分析发现,感染沙门氏菌引起plekhm2基因表达水平升高,plekhm2编码1 016个氨基酸,存在2个基序(RUN和PH),与鸭(Cairina moschata)序列相似性最高,序列比对发现,plekhm2基因cDNA序列第1 013位为错义突变位点c.1013G>A,(p.Gly338Glu),太行鸡为A或G,Kashmir faverolla鸡为A;组织表达谱显示,plekhm2基因在太行鸡心脏、肝脏、脾脏和肺脏等11个组织均有表达;与未感染组相比,感染沙门氏菌后太行鸡肝脏中的plekhm2基因相对表达水平极显著升高(P＜0.01),脾脏中的表达水平显著升高(P＜0.05).plekhm2基因在太行鸡多个组织中均有表达,沙门氏菌感染引起plekhm2基因表达水平升高,本研究为揭示plekhm2基因在沙门氏菌感染中的功能提供了参考.
Chicken salmonellosis is a highly infectious bacterial disease caused by Salmonella infection of chicken(Gallus domesticus).The pleckstrin homology and RUN domain containing M2(plekhm2)gene acts as a target of the Salmonella effector protein SifA and plays a role in Salmonella infection by downregulating the recruitment of kinesin.The aim of this study was to analyze the sequence characteristics of the chicken plekhm2 gene and to investigate its tissue expression profile and the expression levels in the livers and spleens of Taihang chickens infected and uninfected with Salmonella.Differentially expressed genes were screened based on spleen transcriptome data from Salmonella-infected and uninfected chickens,online tools and software were used for bioinformatic analysis of plekhm2 gene,and qPCR was used to detect the expression levels of plekhm2 gene in different tissues of Taihang chickens,and a Taihang chicken Salmonella infection model was constructed to study the changes of its expression in the chicken liver and spleen.The results showed that the expression level of plekhm2 gene was elevated in the spleen-based transcriptome infected with Salmonella,which encoded 1 016 amino acids,with 2 motifs(RUN and PH),and the highest similarity of sequence with duck(Cairina moschata).Sequence comparison revealed a missense mutation site c.1013G>A at position 1 013 of the cDNA sequence of plekhm2 gene(p.Gly338Glu),A for Taihang chicken or G for Kashmir faverolla chicken;Tissue expression profiles showed that plekhm2 gene was expressed in 11 tissues of Taihang chickens,including heart,liver,spleen and lung.Compared with the uninfected group,the relative expression levels of plekhm2 gene in the liver(P＜0.01)and spleen(P＜0.05)of Taihang chickens were significantly higher after Salmonella infection.plekhm2 gene was expressed in several tissues of Taihang chickens,and Salmonella infection caused an increase in the expression level of plekhm2 gene.The present study provides a reference to reveal the function of plekhm2 gene in Salmonella infection.
Chicken salmonellosis is a highly infectious bacterial disease caused by Salmonella infection of chicken(Gallus domesticus).The pleckstrin homology and RUN domain containing M2(plekhm2)gene acts as a target of the Salmonella effector protein SifA and plays a role in Salmonella infection by downregulating the recruitment of kinesin.The aim of this study was to analyze the sequence characteristics of the chicken plekhm2 gene and to investigate its tissue expression profile and the expression levels in the livers and spleens of Taihang chickens infected and uninfected with Salmonella.Differentially expressed genes were screened based on spleen transcriptome data from Salmonella-infected and uninfected chickens,online tools and software were used for bioinformatic analysis of plekhm2 gene,and qPCR was used to detect the expression levels of plekhm2 gene in different tissues of Taihang chickens,and a Taihang chicken Salmonella infection model was constructed to study the changes of its expression in the chicken liver and spleen.The results showed that the expression level of plekhm2 gene was elevated in the spleen-based transcriptome infected with Salmonella,which encoded 1 016 amino acids,with 2 motifs(RUN and PH),and the highest similarity of sequence with duck(Cairina moschata).Sequence comparison revealed a missense mutation site c.1013G>A at position 1 013 of the cDNA sequence of plekhm2 gene(p.Gly338Glu),A for Taihang chicken or G for Kashmir faverolla chicken;Tissue expression profiles showed that plekhm2 gene was expressed in 11 tissues of Taihang chickens,including heart,liver,spleen and lung.Compared with the uninfected group,the relative expression levels of plekhm2 gene in the liver(P＜0.01)and spleen(P＜0.05)of Taihang chickens were significantly higher after Salmonella infection.plekhm2 gene was expressed in several tissues of Taihang chickens,and Salmonella infection caused an increase in the expression level of plekhm2 gene.The present study provides a reference to reveal the function of plekhm2 gene in Salmonella infection.