BmNPV半胱氨酸蛋白酶基因的核苷酸序列研究

对家蚕核型多角体病毒苏州株（BmNPVsu）光胱氨酸蛋白酶基因（CP）的序列分析表明,该基因读码框为972个核苷酸,编码323个氨基酸。同源性分析表明,BmNPVsu的CP与首蓿银纹夜蛾核型多角体病毒（AcNPV）、美国白蛾核型多角体病毒（HcNPV）、云杉卷叶蛾核型多角体病毒（CfNPV）、黄杉毒蛾核型多角体病毒（OpNPV）、舞毒蛾核型多角体病毒（Ld-NPV）在DNA水平上的同源性分别为96.5％、76.2％、74.9％、72.7％、62.9％;在氨基酸水平上的同源性分别为96.9％、77.1％、79.3％、77.1％、65.6％。BmNPV CP的氨基酸序列与不同来源的木瓜蛋白酶超家族的CP也具有较高的同源性,特别与 Trpanosoma brucei的CP具有较高的一致性,达32％。在组织蛋白酶B、H、L、S以及木瓜蛋白酶的36个保守氨基酸残基中有31种出现在BmNPVsu的CP中,BmNPVsu CP同其它杆状病毒CP一样,可看作木瓜蛋白酶超家族成员。     

饲粮添加过瘤胃半胱氨酸对育成期梅花鹿生长性能、营养物质表观消化率和血清生化指标的影响

本试验旨在研究饲粮添加过瘤胃半胱氨酸对育成期梅花鹿生长性能、营养物质表观消化率和血清生化指标的影响。采用单因素随机区组试验设计,将12头7月龄雄性梅花鹿仔鹿分为4组(每组3个重复,每个重复1只),组间体重差异不显著。对照组(Ⅰ组)饲喂基础饲粮,试验组分别饲喂额外添加1.25(Ⅱ组)、2.50(Ⅲ组)和5.00 g·kg^-1(Ⅳ组)过瘤胃半胱氨酸(有效成分51.70%)的试验饲粮。试验预饲期15 d,正试期112 d。结果表明,1)与Ⅰ组相比,添加过瘤胃半胱氨酸Ⅲ组总增重和平均日增重显著提高(P<0.05),Ⅲ组料重比显著降低(P<0.05);2)与Ⅰ组相比,添加过瘤胃半胱氨酸各组粗蛋白质、钙和磷表观消化率显著提高(P<0.05),Ⅲ组干物质和有机物表观消化率显著提高(P<0.05);3)与Ⅰ组相比,添加过瘤胃半胱氨酸Ⅱ组和Ⅲ组甘氨酸表观消化率显著提高(P<0.05),各组半胱氨酸、蛋氨酸、丝氨酸、赖氨酸、苏氨酸、缬氨酸、异亮氨酸、亮氨酸、精氨酸、苯丙氨酸、组氨酸、天冬氨酸、酪氨酸、谷氨酸、丙氨酸和脯氨酸表观消化率显著提高(P<0...     

昆虫取食和剪叶刺激对油松针叶内部分防御物质的诱导效应

为了研究自然条件下昆虫取食及剪叶刺激对油松诱导抗性的影响,本试验应用香草醛—盐酸法,亚硝酸钠—硝酸铝比色法以及紫外分光光度法,分析了剪叶和不同程度油松毛虫取食处理后,混交林和纯林中油松针叶内部分次生代谢物和蛋白酶抑制剂活性的动态变化。结果表明:剪叶及不同程度油松毛虫取食能够诱导缩合单宁、黄酮含量和胰蛋白酶抑制剂、胰凝乳蛋白酶抑制剂活性增加。与对照相比,胰凝乳蛋白酶抑制剂在混交林中的活性比纯林更为明显。说明剪叶及昆虫取食可诱导增加油松针叶内的缩合单宁和黄酮含量、提高胰蛋白酶抑制剂和胰凝乳蛋白酶抑制剂的活性,进而增强油松的抗虫性,林分类型可在一定程度上影响油松诱导防御物质的变化。     

淀粉共生蛋白对淀粉水解的影响

以小麦面粉为原料,研究了淀粉共生蛋白对淀粉酶解性质的影响。对洗淀粉过程中影响蛋白含量的条件进行了单因素试验,用SDS溶液和蛋白酶进一步除去淀粉中的蛋白质成分,然后对制得的含氮量不同的淀粉样品进行酶解。试验结果表明,蛋白酶能有效地去除淀粉共生蛋白,而淀粉共生蛋白有助于淀粉的水解。     

Specificity of polycllonal and monoclonal antibodies for the identification of Xanthomonas campestris pv. campestris

Polyclonal and monoclonal antibodies (PCAs and MCAs), produced to whole cells and flagellar extracts ofXanthomonas campestris pv.campestris (Xcc), respectively, were tested for specificity. In immunofluorescence microscopy (IF) the three PCAs tested, reacted at low dilutions with all Xcc strains, some other xanthomonads and non-xanthomonads. At higher dilutions most cross-reactivity with non-xanthomonad strains disappeared. However, the cross-reactivity with strains ofX. c. pv.vesicatoria (Xcv),X. c. pv.amoraciae (Xca) andX. c. pv.phaseoli var. fuscans (Xcpf) remained.Six MCA-producing cell clones viz. 20H6, 2F4, 18G12, 10C5, 17C12 and 16B5 were selected for specificity tests with an enzyme immunoassay (EIA), IF and a dot-blot immunoassay (DBI). None of the MCAs reacted with all Xcc strains in IF and EIA. In DBI, only MCAs 17C12 and 16B5 reacted with all Xcc strains. All six MCAs tested, cross-reacted in one of either tests with other pathovars ofX. campestris, such as Xcv or Xca. The MCAs were also tested in immunoblotting experiments using total bacterial extracts, cell envelope and flagellar extracts. MCAs 20H6, 2F4, 18G12 and 10C5 reacted with the lipopolysaccharide (LPS) of Xcc. MCAs 16B5 and 17C12 reacted with a 39 kilodalton and a 29 kilodalton protein, respectively.It is concluded that the PCAs and MCAs discussed in this study may be used for routine identification and differentiation of (a group of) Xcc strains. The significance of the cross-reactions with other pathovars ofX. campestris needs to be determined by testing seed lots.     

Sulphur accumulation after Verticillium dahliae infection of two pepper cultivars differing in degree of resistance

Elemental sulphur levels, sulphur localization in stems, and levels of sulphate, glutathione and cysteine were studied in pepper ( Capsicum annuum ) cvs Yolo Wonder (higher resistance) and Luesia (lower resistance) after inoculation with Verticillium dahliae , the cause of vascular wilt. Accumulation of elemental sulphur (S⁰) was first detected 10 days after inoculation in Yolo Wonder (mean S⁰ level 7·3  µ g g⁻¹ DW), and 15 days after inoculation in Luesia (mean S⁰ level 3·3  µ g g⁻¹ DW). The maximum level was reached 21 days after inoculation in Yolo Wonder (14·1  µ g g⁻¹ DW). In control plants, elemental sulphur was not detected. SEM-EDX (scanning electron microscopy-energy dispersive X-ray microanalysis) indicated that the sulphur was not restricted to a specific location, but was dispersed throughout the vascular tissue. Sulphate levels showed a decline at the end of the experiment in inoculated plants, possibly related to the increase in sulphur levels seen in the two cultivars. The differences in sulphate levels between the two cultivars may be due to faster sulphate breakdown in cv. Yolo Wonder.     

哈茨木霉天冬氨酸蛋白酶基因SA76的3＇RACE扩增和序列分析

由EST获得全长cDNA对于结构基因组学和功能基因组学都是至关重要的,cDNA末端快速扩增技术RACE是该领域中的重要研究方法.利用BD SMART RACE技术扩增编码分泌天冬氨酸蛋白酶SA76基因的3＇末端,将其与哈茨木霉cDNA文库中的SA76基因的EST序列进行序列拼接,获得2019bp的全长cDNA序列,其开放读码框长1593bp,5＇非编码区266bp,3＇非编码区201bp,编码530个氨基酸,有信号肽.哈茨木霉天冬氨酸蛋白酶基因与玉蜀黍赤霉、粗糙脉孢菌、球毛壳菌天冬氨酸蛋白酶基因的同源性分别为53%, 37%, 36%.利用BD SMART RACE技术首次从哈茨木霉中克隆天冬氨酸蛋白酶基因,为验证SA76基因的功能奠定基础,为进一步研究蛋白酶的作用机制及生物防治功能提供依据.     

寄生虫半胱氨酸蛋白酶核酸疫苗的研究进展与应用前景

就寄生虫半胱氨酸蛋白酶的一些生理生化与免疫学性质进行了概括,阐述了该酶与宿主免疫应答之间的作用关系,证明了该酶具有作为研制高效核酸疫苗的条件,同时综述了寄生虫半胱氨酸蛋白酶核酸疫苗的发展以及应用前景。     

许氏平鮋消化道中部分消化酶的研究

对许氏平 (Sebastesschlegeli)消化道内的 5种与消化吸收有关的酶进行分布和活性测定。结果表明 ,(1)肠道各段均具很高碱性磷酸酶活性 ,最适 pH为 9.7。食道和胃几乎没有碱性磷酸酶活性。 (2 )消化道各部位均含酸性磷酸酶 ,肠道活性最高 ,自幽门盲囊起依次递降。食道和胃酸性磷酸酶活性明显低于肠 ,胃和肠酸性磷酸酶的最适 pH均为 5 .3。 (3)肠道中羧酸酯酶活性明显高于胃 ,且 2者羧酸酯酶最适 pH差别较大 ,分别为 8.4和 6 .8。(4)胃蛋白酶活性明显高于肠道 ,胃蛋白酶最适pH为 2 .6 ,肠蛋白酶最适 pH为 9.4。 (5 )胃淀粉酶活性明显低于肠道 ,胃淀粉酶最适 pH为 4 .6 ,肠道淀粉酶最适pH为 7.0。这表明许氏平对蛋白质的初步消化在胃中进行 ,在肠中进一步消化和吸收 ,而对糖类和脂类的消化和吸收主要在肠道进行