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南非2型口蹄疫病毒VP1基因的核苷酸及其氨基酸序列分析 被引量:3
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作者 刘亚丽 丁耀忠 +2 位作者 马小元 张杰 张永光 《黑龙江畜牧兽医》 CAS 北大核心 2016年第5期22-26,共5页
VP1基因的遗传衍化关系是口蹄疫病毒分型的依据,而且许多重要的抗原位点也都位于VP1蛋白上。为了对南非2型口蹄疫病毒(SAT2-FMDV)VP1的编码基因序列及其氨基酸序列进行分析,为南非2型口蹄疫的诊断及疫苗设计提供理论基础,试验从NCBI下... VP1基因的遗传衍化关系是口蹄疫病毒分型的依据,而且许多重要的抗原位点也都位于VP1蛋白上。为了对南非2型口蹄疫病毒(SAT2-FMDV)VP1的编码基因序列及其氨基酸序列进行分析,为南非2型口蹄疫的诊断及疫苗设计提供理论基础,试验从NCBI下载大量的南非1/2/3型口蹄疫病毒(SAT1/2/3-FMDV)的1D2A2B基因片段,通过序列分析选择相似性较低的序列代表SAT1/2/3,将选中的代表序列连接于p UC57载体上进行合成,再对合成的含目的序列的穿刺菌进行划板、挑单克隆、摇菌、提质粒,得到重组质粒初步进行酶切鉴定后进行测序,测序正确后采用DNAStar软件包对这些合成序列进行分析。结果表明:SAT2-FMDV VP1的编码基因核苷酸序列变异度较大;虽然SAT2口蹄疫病毒VP1蛋白上主要抗原位点位置与其他血清型的差异性并不大,但是关键氨基酸已发生变化。 展开更多
关键词 口蹄疫病毒 南非型 VP1基因 序列分析 核苷酸序列 氨基酸序列
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南非Ⅱ型口蹄疫主要抗原表位串联基因的表达及抗原性分析 被引量:1
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作者 林祥梅 邓俊花 +1 位作者 王彩霞 吴绍强 《中国畜牧兽医》 CAS 北大核心 2011年第10期55-58,共4页
在对南非Ⅱ型口蹄疫病毒抗原分析基础上,将已经筛选出的6条抗原性良好的多肽采用柔性linker拼接,人工合成相应核苷酸后连入T载体中。将酶切回收的串联基因(VP1-VP3)克隆于表达载体pGEX-6P-1中,获得重组质粒pGEX-VP1-VP3。该质粒转化于... 在对南非Ⅱ型口蹄疫病毒抗原分析基础上,将已经筛选出的6条抗原性良好的多肽采用柔性linker拼接,人工合成相应核苷酸后连入T载体中。将酶切回收的串联基因(VP1-VP3)克隆于表达载体pGEX-6P-1中,获得重组质粒pGEX-VP1-VP3。该质粒转化于感受态细胞BL21(DE3)plysS中,经IPTG诱导后进行了可溶性分析和Western blotting分析,且对融合蛋白柱上酶切纯化后进行了抗原性分析。重组质粒pGEX-VP1-VP3的PCR和测序结果表明,VP1-VP3串联基因已成功插入pGEX-6p-1载体中;pGEX-VP1-VP3融合蛋白分子质量约为38.3ku,并以包涵体形式存在;Western blotting结果显示,该融合蛋白与南非Ⅱ型FMDV阳性血清能发生特异性反应;酶切纯化后蛋白间接ELISA鉴定结果表明,表达的VP1-VP3蛋白具有良好的免疫原性与反应原性。串联多肽的成功表达,将为南非型口蹄疫血清学检测方法建立奠定基础。 展开更多
关键词 口蹄疫病毒 南非型 合成多肽 表达 抗原性分析
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南非Ⅱ型口蹄疫间接ELISA检测方法的建立
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作者 邓俊花 吴绍强 林祥梅 《中国动物检疫》 CAS 2011年第2期34-36,46,共4页
为满足口岸对南非II型口蹄疫检测、监测的需求,在对南非II型口蹄疫抗原表位分析及相关基因合成的基础上,进行融合蛋白pGEX—VP1-VP3的体外表达,采用柱上酶切方法制备VP1-VP3蛋白。将纯化VP1-VP3抗原包被96微孔板,分别优化抗原包被... 为满足口岸对南非II型口蹄疫检测、监测的需求,在对南非II型口蹄疫抗原表位分析及相关基因合成的基础上,进行融合蛋白pGEX—VP1-VP3的体外表达,采用柱上酶切方法制备VP1-VP3蛋白。将纯化VP1-VP3抗原包被96微孔板,分别优化抗原包被浓度、血清稀释度、酶标二抗稀释度,确定阴性临界值。经反复优化,建立的南非II型间接ELISA检测方法的最佳抗原包被浓度为6.4ug/ml,血清最佳稀释度为1:200,酶标二抗最佳稀释度为1:4000,阴性临界值(0D450)为0.562。检测样品判定标准为:OD450〉0.622为阳性,OD450〈0.502为阴性,0.502≤OD450≤0.622为可疑。特异性试验结果表明,所建立方法与其它血清型口蹄疫间无交叉反应。南非II型口蹄疫间接ELISA检测方法的建立为我国口岸口蹄疫检疫提供了新的查验方法。 展开更多
关键词 口蹄疫病毒 南非II口蹄疫 酶联免疫吸附试验 检测
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南非2型口蹄疫病毒结构蛋白抗原表位研究进展 被引量:3
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作者 李茜 代军飞 +6 位作者 丁耀忠 李国秀 侯谦 Ashenafi Kiros Wubshet 马炳 张永光 张杰 《动物医学进展》 北大核心 2020年第6期81-85,共5页
口蹄疫病毒(FMDV)感染会引起偶蹄动物发生口蹄疫(FMD),产生急性系统性传染性水疱。其分为7个血清型,包括A、O、C型,亚州1型(Asia 1)及南非1、2、3型(SAT1、2、3)。其中,SAT FMDV具有明显的地域性,主要集中在撒哈拉沙漠以南,侵害水牛及... 口蹄疫病毒(FMDV)感染会引起偶蹄动物发生口蹄疫(FMD),产生急性系统性传染性水疱。其分为7个血清型,包括A、O、C型,亚州1型(Asia 1)及南非1、2、3型(SAT1、2、3)。其中,SAT FMDV具有明显的地域性,主要集中在撒哈拉沙漠以南,侵害水牛及野生动物,容易发生抗原变异,疫苗交叉保护效果低。近几年的报告表明,中东地区暴发了SAT2FMDV,跨越了长久以来的地域界限,也对我国的养殖业造成了潜在的威胁。我国在SAT FMDV方面的研究较少,有必要建立特异性和灵敏性高的SAT FMDV监测方法作为战略储备,以防其跨境传播。论文主要介绍了SAT2FMDV结构蛋白抗原表位国内外研究进展,以期为今后研发有关SAT2FMDV战略储备表位疫苗提供理论依据。 展开更多
关键词 南非2 口蹄疫病毒 结构蛋白 抗原表位 疫苗
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南非2型口蹄疫病毒VP1蛋白单抗的制备与特性研究
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作者 朱昱茜 石正旺 +8 位作者 罗俊聪 陈婕 林永玉 席韬 张帆 石鑫泰 郑海学 包世俊 田宏 《病毒学报》 CAS CSCD 北大核心 2024年第4期794-801,共8页
为制备南非2型口蹄疫病毒型特异性单克隆抗体,本研究通过原核系统表达南非2型口蹄疫病毒VP1蛋白,亲和层析的方法纯化目的蛋白。将纯化的VP1蛋白使用弗氏佐剂乳化后,免疫BALB/c小鼠,待加强免疫后取血清效价最稳定的小鼠脾细胞与小鼠骨髓... 为制备南非2型口蹄疫病毒型特异性单克隆抗体,本研究通过原核系统表达南非2型口蹄疫病毒VP1蛋白,亲和层析的方法纯化目的蛋白。将纯化的VP1蛋白使用弗氏佐剂乳化后,免疫BALB/c小鼠,待加强免疫后取血清效价最稳定的小鼠脾细胞与小鼠骨髓瘤SP2/0细胞进行融合,应用间接ELISA筛选针对南非2型VP1蛋白的单克隆抗体。采用特异性试验、叠加试验和抗体亚型鉴定等方法对筛选的单克隆抗体特性进行分析。结果发现,本研究成功表达南非2型VP1蛋白,筛选出2株能稳定分泌特异性针对南非2型VP1蛋白的杂交瘤细胞株(3B2和6F2),其亚型分别为IgG2a和IgG1,叠加试验结果表明2株单抗的叠加系数>40%,说明两株单抗针对不同的抗原位点;单抗的效价可达1∶256000,特异性试验证明2株单抗均不与O型、A型口蹄疫病毒及其他常见猪病病毒产生交叉反应。由此可见,本研究制备的单克隆抗体具有良好的特异性和反应性,为南非2型口蹄疫病毒定型诊断和未来疫苗的研制奠定了基础。 展开更多
关键词 南非2 口蹄疫病毒 VP1蛋白 特异性单克隆抗体
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南非Ⅱ型口蹄疫病毒抗原表位的筛选及抗原性分析 被引量:7
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作者 吴绍强 李雅静 +1 位作者 王彩霞 林祥梅 《中国兽医学报》 CAS CSCD 北大核心 2010年第12期1638-1641,共4页
目前,我国南非Ⅱ型(SATⅡ)口蹄疫病毒(FMDV)的防控形势十分严峻,为了防止SATⅡ型FMD的跨境传入,迫切需要建立其特异的检测方法。本研究以SATⅡ型FMDV结构蛋白氨基酸序列为依据,利用分子生物学软件分析了FMDV结构蛋白VP1~VP3上可能... 目前,我国南非Ⅱ型(SATⅡ)口蹄疫病毒(FMDV)的防控形势十分严峻,为了防止SATⅡ型FMD的跨境传入,迫切需要建立其特异的检测方法。本研究以SATⅡ型FMDV结构蛋白氨基酸序列为依据,利用分子生物学软件分析了FMDV结构蛋白VP1~VP3上可能的抗原表位,并人工合成了8条表位多肽。通过采用SATⅡ型FM-DV阳性血清进行ELISA反应,检测其反应原性;通过采用与载体蛋白偶联的合成肽免疫小鼠,测定小鼠血清中抗体效价,检测合成肽的免疫原性。结果表明,合成的8条多肽均能与SATⅡ型FMDV阳性血清结合,其中的6条多肽免疫小鼠后能产生针对多肽的抗体。本研究为利用串联表位为抗原检测SATⅡ型FMDV抗体方法的建立奠定了基础。 展开更多
关键词 南非口蹄疫病毒 抗原表位 抗原性分析
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Cleaning of South African coal using a compound dry cleaning apparatus 被引量:3
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作者 Li Haibin Luo Zhenfu Zhao Yuemin Wu Wanchang Zhang Cuiyu Dai Ningning 《Mining Science and Technology》 EI CAS 2011年第1期117-121,共5页
The compound dry cleaning principle is briefly described.A beneficiation test on South African coal was conducted using a model compound dry cleaning apparatus.Excellent results were obtained and the optimum operating... The compound dry cleaning principle is briefly described.A beneficiation test on South African coal was conducted using a model compound dry cleaning apparatus.Excellent results were obtained and the optimum operating parameters were determined.They are:an amplitude of 3.0 mm,a motor frequency of 47.5 Hz,an air volume of 50%,a transverse angle of 7°,and a longitudinal angle of-2°.These conditions yield a clean coal containing 11%ash and a coal production of 75%.The organic efficiency,η,is 95.86%.These results show that the South African coal can be separated effectively by compound dry cleaning,which will popularize the compound dry cleaning method. 展开更多
关键词 Compound dry cleaning Model apparatus South African coal
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Influence of Flow Regime on the Vegetation Zonation along Mountain Streams in the Western Cape, South Africa 被引量:1
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作者 Erwin Jacobus Joannes SIEBEN 《Journal of Mountain Science》 SCIE CSCD 2015年第6期1484-1498,共15页
Zonation patterns of riparian vegetation have been sampled and described in mountain streams in two catchments in the Hottentots-Holland Mountains, Western Cape, South Africa. Six main vegetation types that differ in ... Zonation patterns of riparian vegetation have been sampled and described in mountain streams in two catchments in the Hottentots-Holland Mountains, Western Cape, South Africa. Six main vegetation types that differ in structure and species composition, are dominant along these river banks: Aquatic vegetation, Wetbanks, Palmiet, Scrub, Forest and Shrubland(Fynbos). The study aims to correlate the vegetation patterns to flooding patterns, in particular the inundation frequency and stream power. A problem arises: because these catchments are ungauged, like most mountain catchments, with the only weirs at the downstream end of the catchment. Discharge data at the weirs are extrapolated to the sites upstream by multiplication with a factor based on the size of the subcatchment that drains through a sample site. In this way, recurrence intervals for floods in mountain streams are derived. Discharges at sites are also calculated using bed roughness(Manning's n) and slope in straight sections with uniform flow conditions. Stream power is derived from the discharges calculated in this manner. The combination of stream power and recurrence intervals explains the occurrence of most vegetation types occurring on the banks, except for one type: Afromontane Forest. This type is probably more dependent on other factors, such as protection from fire and the depth of the groundwater table. 展开更多
关键词 Hydrology Hydraulics Riparian vegetation Mountain streams Floods Stream power Manning's n.
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A continuously stratified nonlinear model for internal solitary waves in the northern South China Sea 被引量:3
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作者 谢皆烁 蔡树群 何映晖 《Chinese Journal of Oceanology and Limnology》 SCIE CAS CSCD 2010年第5期1040-1048,共9页
A continuously stratified nonlinear model is set up to study the impact of topographical character on the generation of internal solitary waves over a sill by tidal flow.One of the reasons why almost all of the genera... A continuously stratified nonlinear model is set up to study the impact of topographical character on the generation of internal solitary waves over a sill by tidal flow.One of the reasons why almost all of the generated internal solitary waves propagate westward in the northern South China Sea is explained.The model simulations describe the generation and propagation of internal waves well.When the strength of imposed barotropic tides and the water stratification stay unchanged,the steepness of the sill slope can control both(a) whether or not the waves induced over a sill by tidal flow are linear internal waves or nonlinear internal solitary waves,and(b) the amplitude of the internal solitary waves generated.If the steepness of the sill is asymmetric,the nonlinear internal solitary waves may be induced on the steeper side of the sill.These conclusions are supported by a numerical experiment with a monthly-mean stratification and an actual seafloor topography from the Luzon Strait. 展开更多
关键词 internal solitary waves numerical modeling SILL South China Sea
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Institutional Constraints to the Competitiveness of the South African Citrus Exports
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作者 Portia Ndou and Ajuruchukwu Obi 《Journal of Agricultural Science and Technology(A)》 2013年第7期521-530,共10页
This paper examined the dual institutional constraints arising from the market liberalisation of the South African citrus industry and the stringent food safety health and private standards associated with the globali... This paper examined the dual institutional constraints arising from the market liberalisation of the South African citrus industry and the stringent food safety health and private standards associated with the globalisation of the agri-business industry. The aim of this paper was to explore the institutional challenges influencing the competitiveness of the South African citrus industry in the export markets. It also explored the implications of the institutional environment on market access. The logistic regression model was used to analyse the statistical data while simple ranking was used to indicate the new and emerging forces affecting the profitability of the industry in recent years. The results show that access to market information, economies of scale, compliance with strict food safety and quality standards, lack of proper infrastructure including transport and challenges of managing a business within the international business environment exerted significant effects on the competitiveness of the industry as well as the ability of domestic producers to enter the export markets. There is need for government intervention especially aligned to investing in infrastructure. Coupled with the support of private institutions, the government needs to support the exporters and producers in complying with private standards. 展开更多
关键词 Institutional constraints PERFORMANCE COMPETITIVENESS South African citrus industry export markets.
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Using potential distributions to explore environmental correlates of bat species richness in southern Africa: Effects of model selection and taxonomy
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作者 M. Corrie SCHOEMAN F. P. D. (Woody) COTTERILL +1 位作者 Peter J. TAYLOR Ara MONADJEM 《Current Zoology》 SCIE CAS CSCD 2013年第3期279-293,共15页
We tested the prediction that at coarse spatial scales, variables associated with climate, energy, and productivity hy- potheses should be better predictor(s) of bat species richness than those associated with envir... We tested the prediction that at coarse spatial scales, variables associated with climate, energy, and productivity hy- potheses should be better predictor(s) of bat species richness than those associated with environmental heterogeneity. Distribution ranges of 64 bat species were estimated with niche-based models informed by 3629 verified museum specimens. The influence of environmental correlates on bat richness was assessed using ordinary least squares regression (OLS), simultaneous autoregressive models (SAR), conditional autoregressive models (CAR), spatial eigenvector-based filtering models (SEVM), and Classification and Regression Trees (CART). To test the assumption of stationarity, Geographically Weighted Regression (GWR) was used. Bat species richness was highest in the eastern parts of southern Africa, particularly in central Zimbabwe and along the western border of Mozambique. We found support for the predictions of both the habitat heterogeneity and climate/productivity/energy hypothe- ses, and as we expected, support varied among bat families and model selection. Richness patterns and predictors of Miniopteridae and Pteropodidae clearly differed from those of other bat families. Altitude range was the only independent variable that was sig- nificant in all models and it was most often the best predictor of bat richness. Standard coefficients of SAR and CAR models were similar to those of OLS models, while those of SEVM models differed. Although GWR indicated that the assumption of stationa- rity was violated, the CART analysis corroborated the findings of the curve-fitting models. Our results identify where additional data on current species ranges, and future conservation action and ecological work are needed. 展开更多
关键词 CHIROPTERA MACROECOLOGY Niche-based models Spatial models Species richness
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