【作者】 肖文福； 蒋亚明； 粟思源； 刘彬斌； 张智勇； 张宇靖； 万林荣； 张友洪；

【Author】 XIAO Wen-fu;JIANG Ya-ming;SU Si-yuan;LIU Bin-bin;ZHANG Zhi-yong;ZHANG Yu-jing;WAN Lin-rong;ZHANG You-hong;Sericultural Research Institute,Sichuan Academy of Agricultural Sciences;

【机构】 四川省农业科学院蚕业研究所；

【摘要】 为了分析通过转录组学和甲基化组学筛选的候选基因在抗性品系和敏感品系高温处理后的表达特征,通过qRT-PCR分析了候选基因在不同品系中表达特征。结果显示,候选基因BmSAMS基因在家蚕抗高温高湿品系中普遍高量表达,在低抗品系中表达量相对较低。为了分析BmSAMS基因的蛋白水平的潜在功能特征,通过生物信息学分析了该基因结构、表达和定位特征,确定该基因在昆虫中高度保守,具有典型的S-AdoMet结构域,BmSAMS基因在家蚕后部丝腺表皮、中肠、马氏管和脂肪体等抗性相关组织高量表达,在家蚕细胞质、细胞核、线粒体和高尔基体等细胞器中均高量表达。过表达BmSAMS基因,在27℃和35℃处理后,细胞增殖活力都明显上升,可以提高细胞增殖活力以抵抗细胞在极端温度条件下的劣势,使细胞能够稳定存活。通过免疫共沉淀筛选了BmSAMS可能的相互作用蛋白,结果显示BmSAMS与核糖体蛋白S3(Ribosomal ProteinS3,RPS3)、异质核核糖核蛋白K(heterogeneous nuclear ribonucleoprotein K,hnRNPK)、rRNA加工蛋白Ebp2(rRNA processing protein Ebp2)、骨形态发生蛋白-2(bmp-2 protein isoform X1,BMP-2)、核糖体蛋白S3a(Ribosomal Protein S3,RPS3a)和肌动蛋白(Actin)具有潜在的相互作用。我们进一步对BmSAMS基因与它们之间的调控关系进行探究,结果显示,过表达BmSAMS基因后能够显著促进BmPPE、BmActin和BmRPS3基因表达;对BmHNRK、BmRPS3a和BmBMP2基因具有明显的抑制作用。更多还原

【Abstract】 In order to analyze the expression characteristics of candidate genes selected by transcriptomics and methylomics after high temperature treatment in resistant and sensitive strains,the expression characteristics of candidate genes in different strains were analyzed by qRT-PCR.The results showed that the candidate gene BmSAMS was highly expressed in the high temperature and humidity resistant strains,but was relatively low in the low temperature and humidity resistant strains.In order to analyze the potential functional characteristics of BmSAMS gene at protein level,the structure,expression and localization of BmSAMS gene were analyzed by bioinformatics.It was confirmed that BmSAMS gene was highly conserved in insects and had a typical S-AdoMet domain.BmSAMS gene was highly expressed in the epidermis,midgut,malpighian tubule and fat body of posterior silk gland of silkworm,and highly expressed in cytoplasm,nuclear,mitochondria and Golgi apparatus.Overexpression of BmSAMS gene significantly increased the proliferation activity of cells after treatment at 27 ℃and 35 ℃,which could improve the proliferation activity of cells to resist the disadvantage of cells under extreme temperature conditions and make them survive stably.The possible interaction proteins of BmSAMS were screened by immunocoprecipitation.The results showed that BmSAMS potentially interacted with ribosomal protein S3(RPS3),heterogeneous nuclear ribonucleoprotein K(hnRNPK),rRNA processing protein Ebp2,bmp-2 protein isoform X1(BMP-2),ribosomal protein S3(rps3a) and actin.We further explored the relationship between BmSAMS gene and their regulation.The results showed that overexpression of BmSAMS gene could significantly promote the expression of BmPPE,BmActin and BmRPS3 genes,and significantly inhibit BmHNRK,BmRPS3a and Bm BMP2genes.更多还原