JS/1/97/Go株GPMV NP基因的克隆、序列分析

根据GenBank中发表的GPMV SF02株的NP基因的核苷酸序列设计合成一对引物,采用RT-PCR扩增出与预期设计的1470bp大小相符的片断,将此扩增产物克隆入PMD18-T载体,进行序列测定和分析.结果表明:所扩增的NP基因的核苷酸长为1 470bp,共编码490个氨基酸.同源性分析表明:JS/1/97/Go与国内的SF02株的NP基因核苷酸同源性为98.6%,氨基酸同源性为99.6%.由此可见,鹅副粘病毒JS/1/97/Go分离株与SF02株的亲缘关系较近,同属于新城疫Ⅶ型基因.而其与LaSota株的核苷酸同源性为85.2%,氨基酸同源性为90.8%.说明该毒株相对于经典的NDV在NP基因上已发生了较大的变异.     

Effects of Time and Rate of Nitrogen and Phosphorus Application on the Growth and the Seed and Oil Yields of Canola (Brassica napus L.)

A field study was conducted to investigate the influence of variable rates of application of N and P fertilizers in splits at various times on the growth and the seed and oil yields of canola (Brassica napus L.) during 1995–97. Rates of fertilizer application were 0 and 0 (F0), 60 and 0 (F1), 0 and 30 (F2), 60 and 30 (F3), 90 and 60 (F4) and 120 and 90 (F5) kg N ha^?1 and kg P₂O₅ ha^?1. All the P was applied at sowing while N was applied in splits, i.e. all at sowing, half at sowing and half with first irrigation, or half at sowing and half at flowering. The responses of growth, seed yield and components of yield were consistent in both years. Increasing the rate of fertilizer application from F4 (90/60 kg N/P₂O₅ ha^?1) to F5 (120/90 kg N/P₂O₅ ha^?1) increased the leaf area index (LAI) relative to the control and to lower rates of fertilizer application. For both crops, application of 90/60 kg N/P₂O₅ ha^?1 significantly enhanced total dry matter (TDM) and seed yield. Seed yield increased mainly due to a greater number of pods per plant and seeds per seed‐pod. The time of fertilizer application did not significantly affect seed yield or components of yield in either season. Oil yield generally followed seed yield, increasing with increasing rate of fertilizer application up to 90/60 kg N/P₂O₅ ha^?1. The maximum oil contents were obtained from the control. The results show that seed and oil yields of canola were maximized at the F4 (90/60 kg N/P₂O₅ ha^?1) rate of application under the agro‐ecological conditions of Faisalabad, Pakistan.     

抗A型禽流感病毒核蛋白特异性单克隆抗体研究

禽流感（AI）是由A型禽流感病毒（AIV）引起的一种发生于禽类的病毒性传染病。笔者以杂交瘤技术研制抗AIV共同抗原的特异单克隆抗体，旨在建立一种双抗体夹心ELISA方法快速检测AIV，以便为A型AIV的快速诊断技术的研究提供理论依据。     

抗A型禽流感病毒核蛋白特异性单克隆抗体研究

利用禽流感H9亚型病毒(AIV-H9)免疫Balb/c小鼠,取其脾细胞与骨髓瘤细胞进行融合,经免疫荧光试验(IFA)检测,以研制抗禽流感病毒(AIV)单克隆抗体。结果获得了5株特异性抗AIV核蛋白(NP)的单克隆抗体细胞株,分别命名为AIV-NP-2C3、AIV-NP-6A5、AIV-NP-3H9、AIV-NP-7B4、AIV-NP-2H4。这5株单克隆抗体能与所有试验的AIV-H9病毒反应,Western blotting方法鉴定结果表明,单克隆抗体仅识别60 ku的蛋白抗原,而不与新城疫病毒、禽网状内皮组织增殖症病毒、传染性法氏囊病毒等反应。初步应用结果显示,以这些单克隆抗体建立的间接免疫荧光试验或ELISA方法可迅速检测出禽流感病毒,这些单克隆抗体在禽流感的预防监测中将发挥重要的作用。     

新城疫病毒F48E9株NP基因的克隆与序列分析

根据NDV NP蛋白已知基因序列设计合成了一对引物，利用RT-PCR扩增出了F48E9株NP基因1598bp的片段，将该片段克隆到pMD18-T Vector上，并对所得到的重组质粒进行酶切分析、PCR鉴定，结果证明我们得到了这个基因片段的阳性重组子。为了更充分证实所得阳性质粒的可靠性，采用Sanger's双脱氧末端终止法对阳性重组子进行核苷酸序列测定，获得了F48E9株NP基因片段的基因序列，利用DNASIS分析软件，将F48E9株与La Sota株的相应序列进行比较，其核苷酸序列同源性为88.2%。至此，我们获得了F48E9株NP基因的全长克隆。     

猪繁殖与呼吸综合征基因标记疫苗株NP49-ELISA鉴别诊断方法的建立

猪繁殖与呼吸综合征病毒（Porcine reproductive and respiratory syndrome virus,PRRSV）rHN4-△25＋NP49株是新型基因标记弱毒疫苗候选株,为了配合对该基因标记疫苗免疫猪血清抗体的鉴别诊断,本研究以标记基因编码的NP49（新城疫病毒NP蛋白C末端49个氨基酸,即NP49）多肽作为包被抗原,建立标记疫苗免疫猪血清中NP49抗体的ELISA鉴别诊断方法。通过对NP49-ELISA工作条件的优化,结果显示NP49-ELISA的多肽抗原最适包被浓度为500ng／孔,被检血清最佳稀释度为1：40。利用ROC曲线法确定S／P临界值为0．2,该方法批内与批间重复试验的变异系数均小于10％,与几种常见猪病阳性血清无交叉反应,具有较好重复性和特异性。本研究建立的NP49．ELISA鉴别诊断方法为猪繁殖与呼吸综合征新型基因标记疫苗的临床应用提供了有力保障。     

新城疫病毒F48E9株M和NP基因的原核表达

根据新城疫病毒（NDV）F48E9国内标准强毒株编码序列设计了2对特异性引物,分别用于扩增M和NP基因。经序列测定,将其克隆到原核表达载体pET-30a（＋）上,转化E．coliBL21（DE3）,筛选表达NDVF48E9株M蛋白和NP蛋白的菌株。IPTG诱导表达后,SDS-PAGE分析结果表明,NDVM蛋白在E．coliBL21（DE3）内以可溶形式存在,而NP蛋白以包涵体的形式表达。用纯化的M和NP蛋白免疫鸡制备超免疫血清,进行Western-blot分析;结果,它们可以与真核表达系统表达的M和NP蛋白以及NDV发生特异性反应,表明,原核表达系统表达的NDVM和NP蛋白具有良好的免疫原性和反应原性。     

Effect of NP9 on gene expression profile of NPC cell line CNE1

AIM: To identify the gene expression profiles of CNE1 cells stably transfected with NP9 expressing plasmid, and to explore the potential molecular function of NP9 gene.METHODS: CNE1 cells stably expressing NP9 protein and CNE1 cells transfected with empty vector were used as test and control. Differentially expressed genes were screened with high- throughout human genome array. Differential expression of 6 genes was analyzed with quantitative RT-PCR. RESULTS: Of all the 14 500 human genes in array, 266 genes were revealed differential expression between test and control , of which 82 genes (RA>1)were up-regulated in test and 184 genes (RA<1) were down-regulated. 34 genes and 75 genes were found distinctively up-regulation (RA>1.5) and down-regulation (RA<1.5),respectively.CONCLUSION: NP9 expression in CNE1 cells leads to changes of some genes involved in regulation of cell cycle, cell proliferation and differentiation, cell signal transduction, cell adhesion. Some new clues may be provided for further studying the potential function and molecular mechanism of NP9 gene.     

两株A型禽流感病毒NP基因的PCR扩增及序列分析

研究选择 AIV NP基因为目的基因 ,设计了一对特异性引物 ,筛选出最佳的 RT-PCR扩增条件 ,特异性地扩增出了两株 A型禽流感病毒的部分 NP基因片段 ,并对其进行了序列分析 ,结果表明 ,两株来源于不同禽类的 AIV的NP基因同源率高达 92 %。AIV NP基因结构具有高度保守性     

新疆不同类型春小麦品种花后干物质与氮磷的积累及转运规律研究

[目的]在自然生态条件下,研究不同品质类型春小麦品种干物质及氮磷的积累和转运规律.[方法]选用新疆主栽的8个春小麦品种,测定其花后干物质和氮磷的积累量,对其积累和转运规律进行研究.[结果]不同类型品种花后干物质与氮磷的积累动态均呈上升的趋势,成熟期干物质与氮磷的积累量强筋品种最高,其次为中筋品种,中强筋品种较低;从干物质与氮磷的转运情况来看,花前干物质转运量和干物质转运率由大到小依次为中强筋＞中筋＞强筋,花后干物质转运量为中强筋＞强筋＞中筋,花前于物质转运对籽粒的贡献率依次为强筋＞中筋＞中强筋.三种类型春小麦品种氮磷的积累与转运状况基本一致,氮磷花后积累量和花前氮磷转移量均为强筋＞中筋＞中强筋.[结论]不同类型春小麦品种之间干物质与氮磷的积累及转运存在一定程度的差异.