MSTN基因编辑牛肌肉转录组测序及生物信息学分析     

苗曼宁  郭益文  胡德宝  曾雨晗  李新  张林林  丁向彬  郭宏 《中国畜牧兽医》2021,48(7):2271-2281

为探究肌生长抑制素（myostatin,MSTN）基因对牛肌肉发育的具体调控机制,本研究选取同一牛场健康鲁西黄牛10头,其中通过转基因技术得到的基因编辑牛MSTN^-/-和同种非转基因野生型牛各5头。分别采集两组牛腿臀肌肉样品,利用IIlumina HiSeq高通量测序技术进行转录组测序分析,通过生物信息学方法比较两组样本间的差异表达基因,并进行GO和KEGG富集分析,最后利用实时荧光定量PCR验证转录组测序数据。结果显示,基因编辑型牛和野生型牛之间共检测到18 071个基因。在log₂|FoldChange|≥ 1.48条件下,筛选出406个差异表达基因,其中347个显著上调,59个显著下调。GO功能富集分析显示,MSTN基因编辑后显著影响915个功能类别（P<0.05）,差异基因主要参与结合、生物系统调节、免疫系统等相关功能。KEGG通路富集分析结果共涉及211个通路,差异基因主要富集在细胞黏附分子、趋化因子信号通路、细胞因子互作等信号通路上,进一步从中筛选出可能参与细胞生长、肌肉发育的差异基因（CD14、KIT、CSF1R、FBP1、DUSP4、ULBP21、PRKCB、SPN、CHAD、SRC）。实时荧光定量PCR检测结果显示,所选差异基因表达水平与转录组表达水平一致,证明测序结果的可靠性。本研究结果表明,MSTN基因发挥作用后可以介导多个下游基因表达,从而影响相关信号通路及生物学过程;同时,所筛选出的差异表达基因可作为进一步研究骨骼肌调控机制的候选靶标。  相似文献   

MSTN基因对牛肌动蛋白细胞骨架调节通路的影响     

盛辉  郭益文  张林林  谭皓云  胡德宝  李新  丁向彬  郭宏 《中国畜牧兽医》2021,48(10):3554-3564

为探究肌肉生长抑制素（myostatin,MSTN）对牛骨骼肌生长发育的作用机制,本研究前期利用定量蛋白质组学与磷酸化蛋白质组学分析野生型蒙古牛（MG.WT）和MSTN^+/-蒙古牛（MG.MSTN^+/-）腿臀肌肌肉组织中蛋白质水平和磷酸化修饰水平的差异变化,使用已建立的牛骨骼肌卫星细胞体外诱导成肌分化模型,检测设计合成的MSTN siRNA （si-MSTN）干扰效果;采用实时荧光定量PCR和Western blotting方法检测转染si-MSTN的增殖期（GM）和分化第3天（DM3）牛骨骼肌卫星细胞中肌动蛋白细胞骨架调节通路相关基因的mRNA和蛋白水平的表达变化,研究敲低MSTN表达对肌动蛋白细胞骨架调节通路的影响。结果显示,在MSTN^+/-蒙古牛肌肉组织中共鉴定到16个肌动蛋白细胞骨架调节通路相关基因表达丰度上调;转染si-MSTN细胞中的MSTN表达水平极显著降低（P<0.01）;在转染si-MSTN的GM期牛骨骼肌卫星细胞中,肌动蛋白细胞骨架调节通路相关基因ENAH、ACTN4和Cdc42的mRNA水平均显著升高（P<0.05）,PFN1、RhoA和ACTN4的蛋白水平均显著或极显著升高（P<0.05;P<0.01）;在转染si-MSTN的DM3牛骨骼肌卫星细胞中,ENAH、CFL1、SCIN和Cdc42基因mRNA水平均显著升高（P<0.05）,RhoA基因mRNA水平极显著升高（P<0.01）,PFN1和ACTN4的蛋白水平均显著升高（P<0.05）。结果表明,干扰MSTN可以促进肌动蛋白细胞骨架调节通路相关基因的表达,探明了MSTN可能通过介导肌动蛋白细胞骨架调节通路影响牛骨骼肌卫星细胞增殖和成肌分化的分子机制,为进一步研究MSTN对牛成肌分化的调控机制提供参考。  相似文献   

Analysis on Differential Expression Genes in Porcine Aortic Vascular Endothelial Cells Induced by Apolipoprotein C Ⅲ     

YUE Yuan  CHEN Hong-yan  WANG Jia-wei  XU Ming-qiang  DING Yu  JIANG Hao  GAO Yan  ZHANG Jia-bao  YAN Shou-qing 《中国畜牧兽医》2016,43(3):585-591

The aim of this study was to investigate the differential expression genes induced by ApoCⅢ,and study the function of ApoCⅢ.Porcine aortic vascular endothelial cells were successfully isolated using enzyme digestion,and then screened the differential expression genes induced by ApoCⅢ using the Solexa high-throughput sequencing technology.The results showed 647 differential expression genes,including 390 up-regulated genes and 257 down-regulated genes.The qRT-PCR results verified that the gene expression results from Solexa sequencing data were reliable.GO and Pathway analysis showed that the function of differential expression genes were related to immune response,cell apoptosis and death.These findings suggested that ApoCⅢ affected the physiological function of porcine aortic endothelial cells by the molecular pathways of inflammation,cell adhesion and apoptosis,which provided a theoretical basis for further understanding the molecular mechanisms of atherosclerosis caused by ApoCⅢ.  相似文献   

Prokaryotic Expression and Antigenicity Analysis of Porcine Japenese Encephalitis Virus Envelope Protein Domain Ⅲ     

HOU Hua-yan  DING Ling-ling  LV Mao-jie  YANG Bao-shou 《中国畜牧兽医》2016,43(4):934-939

In order to analyze the antigenicity of porcine Japanese encephalitis virus (JEV) E protein domain Ⅲ, which was expressed by pET-28a vector with His-tag and purified through Ni-NTA, the BALB/c mice were immunized with the purified protein.We identified the antigenicity of domain Ⅲ of E protein and the anti-mice and anti-porcine JEV E Ⅲ protein specific antibody titers by SDS-PAGE, Western blotting, indirect ELISA and IFA.SDS-PAGE results showed the expressed target protein existed mainly in the form of inclusion body.Western blotting, ELISA test results showed that the protein had good reactivity with anti-serum.The mice immunized with the purified JEV E Ⅲ protein generated 1×10⁵ anti-JEV E Ⅲ protein specific antibody titers by ELISA, and the porcine immunized with the porcine JEV generated 5.1×10⁴ anti-JEV specific antibody titers.The IFA results showed that JEV E Ⅲ protein anti-serum could identify JEV antigen.The above results showed that the recombinant JEV E Ⅲ had good antigenicity.These results provided important basis for development of diagnostic antigen for JEV.  相似文献   

Screening of Probiotics Inhibiting E. coli K88 in vitro     

WANG Lei  LV Yang  ZHANG Yue  XIE Jia-qian  ZHAO Di  YAN Li-qiong  MEI Hui-min  WANG Lei  YI Dan  CHEN Hong-bo  DING Bin-ying  HOU Yong-qing  WU Tao 《中国畜牧兽医》2016,43(5):1355-1360

The probiotics tested in the experiment were isolated from the intestinal of weaning piglets.The isolated probiotics and E.coli K88 were inoculated into the culture of intestinal porcine epithelial cell-1 (IPEC-1).The activity of lactate dehydrogenase (LDH) in the supernatant was measured after incubating for 2.5 h.At the same time, the probiotics and E.coli K88 were co-cultured in vitro, the number of E.coli K88 was counted and the probiotics which could be resistant to the E.coli K88 were selected 2.5 h later.The results showed that the Lactobacillus casei and Bacillus coagulans could significantly reduce LDH activity (P<0.05), decrease the damage of E.coli K88; Bacillus coagulans could inhibit the growth of E.coli K88.At the same time, Bacillus coagulans could resist high temperature, acid and bile salt.The results showed that Bacillus coagulans strains had great potential as the application of probiotics strains.The methods could be used as a model of screen probiotics which could inhibit the growth of E.coli K88 in vitro.  相似文献   

日光温室分段智能屋脊通风效果研究     

尹义蕾  郑禾  李明  何芬  丁小明  周长吉 《北方园艺》2016,(2):39-42

针对现有日光温室人工通风方式劳动强度大、室内气温分布不均匀等问题,在温室内安装2台或3台智能型屋脊通风设备对温室进行2段或3段智能通风,并在北京地区冬季日光温室与人工通风控制进行了对比。结果表明:在保温被揭开期间,当采用3段智能通风时,温室内空气温度最大值与最小值之差为(1.1±0.5)℃;而2段智能控制和人工控制分别为(2.3±1.1)℃和(3.8±1.3)℃。此外,应用该智能屋脊通风技术,还能将667m2均效益提高0.97万元。因此,智能型屋脊通风设备可显著改善温室内温度分布的均匀性,提高种植的经济收益。  相似文献   

苏北沿海滩涂马齿苋高效栽培技术     

刘冲  洪立洲  丁海荣  朱小梅  邢锦城  赵宝泉 《北方园艺》2016,(1):42-44

根据江苏苏北沿海滩涂地区的气候、土壤等因素,结合几年的田间试验,总结了江苏苏北沿海滩涂地区马齿苋高产栽培的适宜播种量、施肥、病虫害防治、采收等栽培管理措施。  相似文献   

不同性能薄膜对草莓气调保鲜效果的影响   总被引：2，自引：0，他引：2  

丁华  王建清  王玉峰  董婧 《北方园艺》2016,(18):126-130

以PE/OPP/PE复合膜、PE膜及电晕处理PE膜为试材,采用与空气相同(21%O_2+0.9%CO2)的初始气体比例被动气调包装件,研究不同透气、透湿性能的薄膜对草莓气调保鲜效果的影响。结果表明:在室温(15±2)℃、相对湿度33%~35%的条件下,在草莓6d贮存期内,PE/OPP/PE复合膜保持了草莓较高的感官评价和硬度,电晕处理PE膜更有利于延缓草莓可溶性固形物含量的下降及pH的升高。故PE/OPP/PE复合膜和电晕处理PE膜对草莓的保鲜效果优于PE膜。  相似文献   

福建三明地区柑橘病毒类病原种类的鉴定及检测     

张洁#  王新#  马崇欢  李丁山  刘国坤  丁新伦  吴祖建 《植物保护》2023,49(3):266-271

为了明确福建省三明地区柑橘病毒类病原(病毒和类病毒)种类,利用RT-PCR技术对其进行了鉴定和检测,并对其检出率进行了分析。结果表明,从207份柑橘叶片样品中检出柑橘衰退病毒(citrus tristeza virus, CTV)、柑橘黄化脉明病毒(citrus yellow vein clearing virus, CYVCV)、柑橘叶斑驳病毒(citrus leaf blotch virus, CLBV)和蚜虫致死麻痹病毒(aphid lethal paralysis virus, ALPV)等4种病毒以及柑橘曲叶类病毒(citrus bent leaf viroid, CBLVd)、啤酒花矮化类病毒(hop stunt viroid, HSVd)、柑橘矮化类病毒(citrus dwarfing viroid, CDVd)、柑橘类病毒Ⅴ(citrus viroidⅤ, CVdⅤ)和柑橘类病毒Ⅵ(citrus viroidⅥ, CVdⅥ)等5种类病毒。其中,CTV、CYVCV、CLBV和ALPV的检出率分别是71.01%、66.67%、0.97%和6.28%,CBLVd、HSVd、C...  相似文献   

基于ISSR分子标记的紫云英种质资源遗传多样性及结构分析     

张梦  史鹏飞  刘春增  李本银  丁丽  张琳  郑春风  张成兰  何春梅  施海帆  吕玉虎  常单娜  曹卫东 《草地学报》2023,31(1):50-60

紫云英(Astragalus sinicus L.)是我国重要的稻田绿肥作物,为揭示不同地区种质资源遗传特征,本研究利用ISSR分子标记对77份紫云英种质资源进行了遗传多样性及结构分析。研究发现,77份供试紫云英种质资源遗传相似性系数为0.567 7～0.868 6,可划分为6个聚类。不同地区群体间分化系数为0.136 3,达到极显著水平(P<0.001),其中种群间变异占比13.9%,种群内变异占比86.1%。不同ISSR引物扩增信息鉴别紫云英资源的能力存在差异,构建了以P809第3,8,9,10,12,15扩增位点以及P886第1,3,5,9,10,11扩增位点为特征的指纹图谱,能有效区分77份供试紫云英资源。以资源所在省份为单元进行群体分析,供试紫云英资源遗传结构呈现分化特征,河南-安徽-江苏-上海资源遗传结构相似,构成北方亚群;四川-广西-湖南-江西资源相似,构成西南方亚群;福建种质资源则呈现混合型特征。本研究结果可为我国紫云英种质资源的科学利用及新品种选育提供依据。  相似文献