巨型艾美耳球虫偏菱形样蛋白EmRP免疫原性分析

在前期研究中,从巨型艾美耳球虫(Eimeria maxima)cDNA文库中筛选到了免疫保护性抗原偏菱形样蛋白EmRP,本研究为揭示该抗原的免疫保护机理,进一步检测了其免疫原性。以E.maxima卵囊cDNA为模板,用RT-PCR技术扩增EmRP,并构建真核表达质粒pVAX1-EmRP,将其经腿部肌肉注射2周龄的雏鸡,3周龄加强免疫。分别于首次免疫后和加强免疫后1周,采集血清,用ELISA方法检测血清特异性IgG水平,用荧光定量PCR方法(qPCR)检测细胞因子水平,用流式细胞术检测CD4^+T淋巴细胞和CD8^+T淋巴细胞的比例,分析EmRP诱导的免疫反应。序列分析表明,EmRP含有偏菱形样蛋白超家族成员保守区域,为非跨膜蛋白,分子量约为28.4 kD。真核表达质粒pVAX1-EmRP免疫鸡后,与对照组相比,鸡体IFN-γ、IL-2、IL-10和IL-17等细胞因子转录水平显著提升;CD8^+和CD4^+ T细胞比例显著提高,而血清特异性IgG水平与对照组差异不显著。结果表明,EmRP诱导的免疫保护作用主要是由其诱导的细胞免疫反应实现的,可以作为研制E.maxima新型疫苗的候选抗原。     

不同外源激素对糜子成穗及产量的影响

研究不同种类外源激素对糜子成穗及产量的影响。用5种外源激素在糜子3~4叶期喷施,连续处理2次,喷清水为对照。记录糜子苗期基本苗数、拔节期总茎数、成熟期穗数等性状,用统计软件SPSS 17.0进行统计分析。结果表明,施用脱落酸（ABA）能够显著降低糜子的分蘖数,施用赤霉素（GA₃）、吲哚乙酸（IAA）、矮壮素（CCC）也能减少糜子分蘖数,但是与对照差异不显著,施用多效唑（PP₃₃₃）对糜子分蘖数没有明显影响;施用GA₃、IAA、CCC能够增加分蘖穗数,而ABA、PP₃₃₃降低分蘖穗数;各处理分蘖成穗率都较对照高,ABA处理分蘖成穗率为最高达到25.20%;ABA处理主茎穗占比较对照增加,分蘖穗占比降低;GA₃、IAA、CCC处理主茎穗占比相对于对照降低,分蘖穗占比增加,PP₃₃₃处理与对照相近;5种外源激素对糜子产量的影响差异显著,施用ABA产量最高,可能与主茎穗占比高和分蘖成穗率高有关系。在糜子3~4叶期喷施5种外源激素,对糜子有显著作用的是ABA,主要表现在ABA能够降低糜子分蘖数,保证了主茎的成穗数量,增加了分蘖成穗率,从而提高了糜子产量。     

山羊FGF1O基因的克隆及其表达分析

成纤维细胞生长因子10(fibroblast growth factor 10,FGF10)是一种极其重要的生长因子,能促进小鼠(Mus musculus)和人(Homo sapiens)前体脂肪细胞的分化.为了获得山羊(Capra hircus)FGF10基因序列,研究其组织表达特性,确定其在肌内前体脂肪细胞分化过程中的表达模式,并分析FGF10mRNA表达水平与肌内脂肪沉积的关系,本研究以简州大耳羊(C.hircus)为实验动物,采用反转录聚合酶链式反应(RT-PCR)技术克隆FGF10基因,采用Ⅱ型胶原酶消化获得山羊原代肌内前体脂肪细胞,利用实时荧光定量PCR(qPCR)技术检测FGF10在成年山羊不同组织中的表达差异及其在肌内前体脂肪细胞分化过程中的表达水平,并将基因表达水平与肌内脂肪(intramuscular fat,IMF)含量进行关联分析.结果显示,克隆得到山羊FGF10基因(GenBank登录号:KT899958)序列1 252 bp,其中开放阅读框642 bp,编码213个氨基酸,与绵羊(Ovis aries)和牛(Bos taurus)的该基因同源性达100％,具有跨膜结构域和信号肽结构.FGF10 在山羊肺脏中表达水平最高,显著高于其他组织(P＜0.05),在脾脏和脂肪中也存在较高水平的表达.FGF10 mRNA在成年羊背最长肌中表达量显著高于羔羊与育成羊(P＜0.05),相关性分析结果显示,FGF10 mRNA表达量与山羊背最长肌IMF含量呈显著正相关(P＜0.05).FGF10 mRNA在肌内前体脂肪细胞中表达水平最低,在诱导分化后2d达到最高.本研究结果为进一步阐明FGF10基因在山羊肌内脂肪沉积中的分子机制提供了理论依据.     

高位池循环海水梭子蟹养殖模式探讨(上)

<正>梭子蟹南方又称白蟹。随着梭子蟹商业化捕捞的加剧,捕捞上来的野生梭子蟹个头越来越小,这表明,该物种已被捕捞过度。传统的梭子蟹养殖模式无法适应高速增长的市场以及日益恶化的环境。因此,探究一种新型的梭子蟹养殖模式迫在眉睫。本研究通过建立高位池循环海水系统,利用生物调控,形成蟹     

南方早稻中后期田管技术意见

<正>当前,南方早稻进入大田生长中后期,其中华南地区进入灌浆期、长江中下游地区进入拔节孕穗期。针对当前早稻群体长势和气候特点,农业部水稻专家指导组和全国农业技术推广服务中心会商提出南方早稻中     

沙门菌CRISPR的结构与功能研究进展

近年发现沙门菌中恒定地存在两个CRISPR位点,沙门菌利用它们不仅能抵御外源质粒和噬菌体的入侵,还能介导自身短期的表型变化及长期亚类的进化。同时由于其结构的多样性,使之广泛应用于沙门菌分型和进化的研究。论文简要综述了沙门菌CRISPR系统的基本结构、作用机制和功能及其在沙门菌分型和进化应用方面的研究进展。     

蛋鸡场球虫病的防治

<正>鸡球虫病是最常见且严重危害蛋鸡养殖的寄生虫病,发生率最高。急性暴发时,会引起鸡的大批死亡,慢性发病时则影响鸡的生长、发育,病愈的成年鸡为带虫者,增重和产蛋能力降低,给养鸡业带来巨大损失。因此,鸡球虫病的防治至关重要,特别是地处高湿的山区,这个问题尤为突出。1病原学病原为原虫中的艾美耳科艾美耳属球虫。该病分为急性型和慢性型二种。2流行病学     

狗狗的耳朵

正狗狗的品种众多,它们的体貌形态亦各有千秋。耳朵,是区分它们体征的其中重要一项。也许有人会认为狗狗的耳朵不是立着就是垂着的,那就错了。形态各异的狗耳朵都有特定的叫法。比如巴哥犬的耳朵有玫瑰耳也有纽扣耳,纽扣耳比玫瑰耳更理想。大致来说,狗狗耳朵类型可以分为7种,另外还有几个特殊款,是专门用来形容特定     

Study on Antiviral Effect of Polysaccharides from Lagenaria siceraria (Molina) Standl.on NDV in vitro

To research on polysaccharides from Lagenaria siceraria (Molina) Standl.against Newcastle disease virus (NDV).Total polysaccharide and four different solvent extractions of Lagenaria siceraria (Molina) Standl.were extracted by the methods of hot water extraction and ethanol precipition in this study.The content of polysaccharide was measured by phenol-sulfuric acid method and infrared spectroscopy method.The safe concentration and growth of chick embryo fibroblast (CEF) were assayed by MTT method,in order to facilitated the comparison under the same level,the safe concentration was united as 78.125 μg/mL.Under the safety range of concentration,detected the block-virus activity,anti-virus activity and virus-killing activity of polysaccharides through the ways of pre-adding polysaccharides,post-adding polysaccharides and adding polysaccharides with NDV.The results showed that the direct inactivation and propagation inhibition activity of total polysaccharide and four different solvent extractions were stronger than anti-absorption function.Anti-virus inhibition rate of 70%,80% gradient alcohol precipitation of polysaccharides from Lagenaria siceraria (Molina) Standl.(LSP₇₀,LSP₈₀) and total polysaccharide (LSP_t)were 40.41%,44.54% and 61.85%,virus-killing inhibition rate were 44.74%,58.76% and 59.38%.LSP₈₀ had the highest virus inhibition rate as 37.14% in the block-virus activity of those five polysaccharides.In summary,70%,80% gradient alcohol precipitation and total polysaccharide in polysaccharides possessed better activity and would be as the materials for further research.     

Effect Comparison of Adenovirus Vector Transfer into Different Buffalo Cells

Buffalo mammary epithelial cell,cumulus cell and fibroblasts were transfected by adenovirus vectors and compared their transfection efficiency.293 cells were transfected with pBHGloxdelE13cre and pDC316-eGFP by liposome,the virus was collected and titer was detected.Buffalo mammary epithelial cell,cumulus cell and fibroblasts were exposed to different multiplicity of infection (MOI) of adenovirus vectors.After 72 h,the cells were observed with inverted fluorescence microscope,and transfection efficiency was calculated.When the MOI was 25,50,100,200 and 400,the transfection efficiency of fibroblasts were 0.7%,7.0%,9.0%,12.5% and 34.0%,the transfection efficiency of cumulus cells were 42.5%,55.3%,57.4%,76.0% and 80.0%,the transfection efficiency of mammary epithelial cells were 88.7%,100%,100%,100% and 100%.The results showed that the transfection efficiency of mammary epithelial cell was the best,followed by cumulus cell,and fibroblast was poor.