单晶6H-SiC经氦离子辐照及退火后的微观组织研究

在400℃下对单晶6H-SiC进行了400keV氦离子辐照,辐照剂量为1×1016He+/cm2,随后在1200和1500℃退火30min。采用透射电子显微镜和扫描电子显微镜对辐照态和退火态SiC进行微观结构观察与分析。结果表明,单晶6H-SiC在400℃经氦离子辐照后,仅观察到由辐照引起的位移损伤带,而未观察到明显尺寸的氦气泡。但经1200℃退火30min后,在辐照损伤区域形成了呈血小板状的气泡簇,其主要分布在（0001）晶面上,少量形成在（1120）晶面。辐照未在6H-SiC表面上形成明显尺寸的缺陷,而经1200℃退火30min后,SiC表面出现大尺寸的起泡和凹坑,进一步提高退火温度至1500℃时,表面起泡和形成凹坑更严重,并产生了大量裂纹。本研究同时对微观结构演化的机理进行了分析与讨论。中国工程物理研究院NPL,CAEP项目资助(2015AB001

香蕉横切薄层切片芽分化的培养技术

以香蕉横切薄层切片为材料,研究了不同因素对薄层切片芽分化的影响。结果表明:利用横切薄层培养技术能更有效、快速地进行香蕉的无性繁殖;TDZ(英文名为Thidiazuron,N-pheny-N'-1,2,3.-thia-diazol-5-ylurea,中文译为苯基噻二唑基脲)对香蕉薄层切片芽的分化有抑制作用,没有得到生长正常的不定芽;横切薄层切片在暗培养且培养温度为30℃时比25℃有较高的不定芽分化率,不定芽分化能力强,丛芽多;蔗糖和AgNO_3对薄层切片芽分化有一定的影响

根癌农杆菌介导的芦荟遗传转化条件的研究

以美国库拉索芦荟 (Aloe .arborescens)的横切薄层切片 (transversethincelllayer,tTCL)作为转化受体 ,通过受体材料对抗生素的敏感性实验和Gus基因瞬时表达率的研究 ,找出了较适合的外植体转化条件。研究表明 :芦荟对头雹霉素 (cefotaxime)和羧苄霉素 (carbenicillin)不敏感 ,而对卡那霉素 (kanamycin)和潮霉素 (hygromycin)敏感 ;用靠近顶芽的材料得到的横切薄层切片芽再生率高 ,有较高的Gus基因瞬时表达率 ;乙酰丁香酮 (acetosyringone)在芦荟转化是不可缺少的 ,对其转化有明显的促进作

Preparation of monoclonal antibodies against 3D protein of EV71 based on HBc particles as expression vector

目的:预测肠道病毒71型(EV71)非结构蛋白3D的表位,以HBc蛋白为载体展示多肽,制备并鉴定抗EV71-3D的特异性单克隆抗体(mAb); 。方法:应用生物信息学方法分析预测出EV71; 3D蛋白亲水性和免疫原性指数较高的多肽片段,并运用HBc颗粒型蛋白载体展示肽段,构建多肽融合蛋白,免疫BALB/c雌鼠,通过杂交瘤技术和亲和层析; 技术制备和纯化抗EV71-3D蛋白的特异性mAb,用间接ELISA、ELISPOT、IFA和IHC对mAb的性质进行初步鉴定。结果:构建表达分别; 嵌合3D蛋白34~ 43位氨基酸残基、61~ 76位氨基酸残基、151~; 164位氨基酸残基的HBc重组蛋白,免疫并经过多轮克隆化筛选,获得抗EV71-3D单克隆抗体3E1,其亚类为IgG2a;免疫荧光试验、ELISP; OT法和免疫组织化学染色结果显示其可与EV71特异性结合。结论:成功制备可特异性识别EV71的单克隆抗体3E1,为病毒的检测及进一步研究3D蛋白; 的功能奠定了基础,同时还验证了生物信息学技术与HBc颗粒型载体展示多肽技术相结合可快速高效地制备单克隆抗体。Objective: To prepare and preliminarily identify the monoclonal; antibodies(mAbs) specifically against 3D protein of Enterovirus; 71(EV71),using bioinformatics to predict the epitopes of 3D,with HBc; protein as a carrier.Methods: Artificial screening of 3D protein epitope; sequences by bioinformatic method,inserted into the major immunodominant; region(MIR) area of Hepatitis B virus core protein(HBc),to construct the; recombinant protein.BALB/c mice were immunized with the recombinant; virus like particles(VLPs),to prepare the mAbs against 3D protein of; EV71.Affinity chromatography technology was used to purify the mAb.The; indirect ELISA,ELISPOT,immunofluorescence and immunohistochemistry; staining methods were used to identify the characteristic of the; mAb.Results: We displayed 3D(aa34-43),3D(aa61-76) and 3D(aa151-164); epitopes by constructing fusion protein using HBc VLPs as a vector,after; hybridization,one positive hybridoma cell line(3E1) was selected by; ELISA.The isotype of 3E1 was IgG2a.The results of immunofluorescence and; immunohistochemistry staining assay showed that the mAb 3E1 could; specifically recognize EV71.Conclusion: The prepared mAb 3E1 can; specifically recognizes the EV71,which laid the foundation for the; detection of virus and further study on 3D protein,and verified the; bioinformatics technology combined with HBc carrier displaying peptides; could prepare mAb quickly and efficiently.国家自然科学基金项

人表皮生长因子(hEGF)基因的合成、鉴定及植物表达质粒的构建

用 PCR方法合成了人表皮生长因子 ( h EGF)基因 ,构建了原核表达质粒 p2 0 T-h EGF,研究了原核表达的重组蛋白 h EGF的生物学活性 ,并构建了植物表达质粒 .研究表明 :无论是融合蛋白 GST-h EGF或纯化的h EGF蛋白 ,都有相当高的生物活性 ,h EGF蛋白对 Hela细胞的增殖有很好的促进作用 ,免疫小鼠亦能产生很高的免疫应答反应

影响根癌农杆菌介导的香蕉遗传转化因素研究

以香蕉 (Musa spp.)的横切薄层切片 (transverse thin cell layer,t TCL)外植体作为转化受体 ,通过对外植体 GUS基因瞬时表达率的研究以及受体材料对抗生素的敏感性实验 ,找出了较适合的外植体转化条件和培养条件。研究表明 :用低代香蕉无菌苗为材料 ,横切薄层切片芽再生率高 ,有较高的 GUS基因瞬时表达率 ;香蕉对头孢霉素 (Cefotaxime)和羧苄霉素 (Carbenicillin)不敏感 ,而对潮霉素(Hygromycin)很敏感 ;菌液的预处理是影响香蕉转化的主要因子 ,重悬液中的蔗糖浓度也是影响转化的因素之一 ,对遗传转化有明显的促进作

芦荟横切薄层培养再生植株(简报)

以芦荟横切薄层在改良MS+6-BA3.5mg/L(单位下同)+IBA0.3培养基上,芽诱导率可达95%以上;丛生芽于改良MS+6-BA 2.5+IBA0.2培养基培养可增殖;在改良MS+NAA0.5培养基上经15～20d可产生健壮根系

Preparation and Application of Soluble Human Squamous Cell Carcinoma Antigen Expressed by Escherichia coli

旨在建立基于大肠杆菌表达系统的高效可溶性表达人鳞状上皮细胞癌抗原(SCCAg)方法,获得具有较好活性的重组SCCAg抗原并应用于建立抗原检测方法; 。基于pGEX-6P-l载体和大肠杆菌E. coli; ER2566菌株开展重组SCCAg抗原可溶性表达纯化方法研究,评价纯化抗原活性,筛选特异性单克隆抗体,初步建立并评价SCCAg抗原检测方法。结果; 显示,pGEX-6P-l载体和E coli; ER2566菌株可用于建立较高效的可溶性表达和纯化SCCAg抗原的方法,获得了具有较高纯度和活性的重组SCCAg抗原,筛选获得特异性单克隆抗体并; 初步建立了 SCCAg管式化学发光检测方法。建立了有效的基于大肠杆菌表达系统的可溶性表达和纯化SCCAg的方法。The aims of this study are to establish a method for efficient soluble; expression of human squamous cell carcinoma antigen(SCCAg ) based on; Escherichia coli expression system and obtain the recombinant SCCAg; antigen in fine activity, then apply it in the detection method; establishment of antigen. The study on the method of soluble expression; and purification of recombinant SCCAg antigen was conducted based on; pGEX-6P-l vector and E. coli ER2566 strain. The activity of the purified; antigen was evaluated by Abbott Kit and the specific monoclonal antibody; was screened by indirect ELISA. It was proved that PGEX-6P-1 vector and; E. coli strain ER2566 could be used to establish efficient soluble; expression and purification method for recombinant SCCAg antigen.; Moreover, the recombinant SCCAg antigen was proved to be in high purity; and activity. Thus,the SCCAg detection method of chemical luminous tube; was established with the specific monoclonal antibodies. In conclusion,; an effective method for the expression and purification of SCCAg, which; is based on the E. coli expression system, is established.国家高技术研究发展计划(863计划

香蕉农杆菌介导高效转化体系

以香蕉 (Musaspp .)的横切薄层切片 (tTCL)外植体作为转化受体 ,研究影响根癌农杆菌转化香蕉的因素。研究表明 ,菌液的预处理和重悬液的pH值是影响香蕉转化的主要因子 ,而乙酰丁香酮 (AS)是香蕉遗传转化中必需的酚类物质 ,80 μmol/L是较理想的浓度 ;感染时间以 8～ 15min ,共培养的时间和温度分别以 4～ 5d及 2 6℃为最佳的条