野生瓣化型雄性不育胡萝卜‘武野-不育’的特征鉴定与分析

胡萝卜雄性不育材料,在胡萝卜育种中具有重要作用。本研究描述了一种从武汉市洪山区野生胡萝卜材料中获得的野生瓣化型胡萝卜雄性不育材料(‘武野-不育’,简称‘Wuye-BY’)的特征。该野生瓣化型雄性不育胡萝卜特征在于:无明显膨大的肉质根,嫩茎、叶片及其叶柄深绿色,几乎无毛,无花瓣和花药。花萼数量8~10个,花丝数量为0~2个,染色体数目为2n=18。双悬果顶端及其花丝顶端具有大量的蜜液,能接受栽培胡萝卜花粉。杂交F1的根大小、根重,胡萝卜素、总糖和维生素C含量,具有明显优势。     

丙型肝炎病毒结构蛋白E1在昆虫细胞中的表达及定位

本文在大肠杆菌中表达了与GST融合无跨膜区的丙型肝炎病毒(Hepatitis C Virus,HCV)E1蛋白,并通过免疫兔制备了兔抗E1的抗血清。然后利用Bac-to-Bac杆状病毒表达系统构建了含有HCV结构蛋白E1基因的重组杆状病毒vAcHCVE1。通过Western blot分析,E1蛋白在Sf9细胞中表达分子量大小为30kDa大于预测的20kDa,表明存在翻译后修饰如糖基化等。通过Confocal显微镜观察当感染48h后E1蛋白定位在细胞质和细胞膜上。     

Effects of long-term fixation on histological quality of undecalcified murine bones embedded in methylmethacrylate

While long-term fixation and storage of specimens is common and useful for many research projects, it is particularly important for space flight investigations where samples may not be returned to Earth for several months (International Space Station) or years (manned mission to Mars). We examined two critical challenges of space flight experimentation: the effect of long-term fixation on the quality of mouse bone preservation and the preservation of antigens and enzymes for both histochemical and immunohistochemical analyses, and how the animal/sample processing affects the preservation. We show that long-term fixation minimally affects standard histological staining, but that enzyme histochemistry and immunolabeling are greatly compromised. Further, we demonstrate that whole animal preservation is not as suitable as whole leg or stripped leg preservation for long-term fixation and all histological analyses. Overall, we recommend whole leg processing for long-term storage of bone specimens in fixative prior to embedding in plastic for histological examination.     

连翘种子挥发油抑真菌及在猪油脂酸败过程中的作用

利用滤纸片法观察不同浓度连翘种子挥发油对啤酒酵母、产黄青霉以及黑曲霉的抑制作用,并测定了其最小杀菌浓度;利用猪油体系对连翘种子挥发油在油脂酸败过程中的作用进行了初步探索。结果表明连翘种子挥发油对这三种真菌均有很好的抑制作用,最小杀菌浓度分别为:啤酒酵母为3.91×10-4mL/100 mL,产黄青霉为7.81×10-4mL/100 mL,黑曲霉为3.13×10-3mL/100 mL;而连翘种子挥发油对猪油具有促进其氧化的作用。     

Fibromodulin-deficient mice display impaired collagen fibrillogenesis in predentin as well as altered dentin mineralization and enamel formation.

To determine the functions of fibromodulin (Fmod), a small leucine-rich keratan sulfate proteoglycan in tooth formation, we investigated the distribution of Fmod in dental tissues by immunohistochemistry and characterized the dental phenotype of 1-day-old Fmod-deficient mice using light and transmission electron microscopy. Immunohistochemistry was also used to compare the relative protein expression of dentin sialoprotein (DSP), dentin matrix protein-1 (DMP 1), bone sialoprotein (BSP), and osteopontin (OPN) between Fmod-deficient mice and wild-type mice. In normal mice and rats, Fmod immunostaining was mostly detected in the distal cell bodies of odontoblasts and in the stratum intermedium and was weaker in odontoblast processes and predentin. The absence of Fmod impaired dentin mineralization, increased the diameter of the collagen fibrils throughout the whole predentin, and delayed enamel formation. Immunohistochemistry provides evidence for compensatory mechanisms in Fmod-deficient mice. Staining for DSP and OPN was decreased in molars, whereas DMP 1 and BSP were enhanced. In the incisors, labeling for DSP, DMP 1, and BSP was strongly increased in the pulp and odontoblasts, whereas OPN staining was decreased. Positive staining was also seen for DMP 1 and BSP in secretory ameloblasts. Together these studies indicate that Fmod restricts collagen fibrillogenesis in predentin while promoting dentin mineralization and the early stages of enamel formation.     

Overcoming the thermodynamic limitation in asymmetric hydrogen transfer reactions catalyzed by whole cells

Whole lyophilized cells of an Escherichia coli overexpressing the alcohol dehydrogenase (ADH-'A') from Rhodococcus ruber DSM 44541 were used for the asymmetric reduction of ketones to secondary alcohols. The recycling of the required nicotinamide cofactor (NADH) was achieved in a coupled-substrate process. In the course of the reaction the ketone is reduced to the alcohol and the hydrogen donor 2-propanol is oxidized to acetone by one enzyme. This leads to a thermodynamic equilibrium between all four components determining the maximum achievable conversion. To overcome this limitation an in situ product removal technique (ISPR) for the application with whole cells was developed. In this method the most volatile compound is separated from the reaction vessel by an air flow resulting in a shift of the equilibrium towards the desired secondary alcohol. The so-called stripping process represents a simple and efficient method to overcome the thermodynamic limitation in biocatalytic reactions. Employing this method, the conversion of selected biotransformations was increased up to completeness.     

芜菁类黄酮3′-羟化酶基因的功能鉴定及启动子初步分析

类黄酮3′-羟化酶（Flavonoid 3′-hydroxylase,F3′H）是细胞色素P450单加氧酶,在花青素合成途径中催化二氢山奈酚生成二氢槲皮素,进而形成矢车菊色素。利用津田芜菁BrF3′H1和赤丸芜菁BrF3′H2基因构建过量表达载体后遗传转化烟草,转基因植株的花色加深。通过染色体步移法克隆了BrF3′H1和BrF3′H2基因上游846和851 bp的启动子序列。生物信息学分析表明,BrF3′H1P和BrF3′H2P均包含TATA box、CAAT box、光调控元件、MRE、ABRE、ATGCAAAT-motif、ERE、O2-site、RY-element、LTR等多个顺式作用元件;二者的核苷酸序列在7个位点存在差异。利用BrF3′H1P和BrF3′H2P序列替换pCAMBIA1301植物表达载体的35S启动子后遗传转化烟草。GUS组织化学染色结果表明,BrF3′H1P和BrF3′H2P序列均能驱动GUS基因表达。通过PCR方法获得了BrF3′H1P和BrF3′H2P的一系列缺失片段,融合GUS基因后转化烟草。染色结果显示,BrF3′H1P和BrF3′H2P系列缺失片段均具有起始GUS基因表达的活性。BrF3′H1和BrF3′H2基因的功能鉴定及启动子的初步分析将为揭示津田芜菁和赤丸芜菁F3′H基因的光诱导表达调控机理奠定研究基础。     

棉花根际亲和性高效促生细菌的分离筛选

为了从棉花根际土壤筛选能与棉花凝集素具有亲和作用的高效促生细菌,以选择性培养基从棉花根部初步筛选具有固氮能力、解磷能力及解钾能力的促生细菌,再以异硫氰酸磺(FITC)标记的棉花凝集素为复筛工具,从棉花根际促生细菌中筛选能与棉花凝集素结合的亲和性菌株,分别挑选2株固氮菌、2株解磷细菌和2株解钾细菌作为微生物肥料接种到棉花根部进行盆栽试验.观察其在根部定殖情况.结果是在选择性平板上有20%～30%的菌株具有凝集素染色阳性.盆栽试验显示,接种的6株亲和性菌株能在棉花根部成功定殖,根际细菌数量约是灭活对照的`0倍.通过初步鉴定,固氮菌株N1111为固氮菌属(Azotobacter),N2121属于德克斯氏菌属(Derxia);解磷菌株P2126属于黄单胞菌属(Xanthomonas),P1108菌株为假单胞菌属(Pseudomonas);解钾菌株K2204和K2116属于芽孢杆菌属(Bacillus).     

一起荷斯坦奶牛乳头瘤病诊断及其病毒基因型分析

牛乳头瘤病毒是一种能够引起牛发生皮肤乳头状瘤、纤维素瘤、膀胱和食道癌的DNA病毒,现已在牛群中广泛传播,对牛养殖业造成了重大经济损失.为了诊断甘肃某荷斯坦奶牛场300多头泌乳期奶牛乳头突发疣状物的病因,本研究采用流行病学调查、临床观察、组织病理学、分子生物学检测方法和基因测序技术,对患有疣状物的奶牛进行综合诊断.结果 表明,乳头患疣状物奶牛的其它部位无类似生长物,无发烧、疼痛等异常临床症状,组织病理学HE检测疣状物呈现角化过度和细胞空泡化现象,这与牛乳头瘤病毒感染的组织病理变化相似,并且用PCR方法获得了牛乳头状瘤病毒L1基因,测序比对结果显示为乳头瘤病毒7型基因,核苷酸同源性达98％以上.因此,本次荷斯坦奶牛乳头突发疣状物为乳头瘤病毒7型感染引起的,这是甘肃地区首次发现该基因型乳头瘤病毒,应引起奶牛场与防疫部门的重视.