H5N1亚型禽流感病毒NS1基因的原核表达及其ELISA检测方法的建立   总被引：4，自引：0，他引：4  

杨涛刘明  张云刘春国  杜绍范 《中国兽医科技》2005,35(8):585-589

采用RT-PCR技术扩增了禽流感病毒（AIV）A／Goose／HLJ／p46／2003（H5N1）的NSl基因，将其克隆于融合蛋白表达载体pMAL-c2X上，转化DH5α大肠埃希氏菌感受态细胞，经BamHⅠ和HindⅢ双酶切鉴定及序列分析，表明筛选到了重组质粒pc2X-NS1。SDS-PAGE电泳结果显示，重组质粒转化TB1大肠埃希氏菌后，经0．3mmol／L的IPTG诱导，融合蛋白MBP-NS1得到大量表达，融合蛋白以可溶形式存在，分子质量约为67ku。Western-blotting检测结果表明，融合蛋白MBP-NS1能够与H5N1亚型AIV活病毒感染康复鸭血清发生特异性反应，而不能够与H5N1亚型AIV灭活疫苗免疫鸭血清发生反应。试验初步建立了以纯化的融合蛋白MBP-NS1为包被抗原的间接ELISA检测方法，为AIV灭活疫苗免疫家禽与AIV感染家禽的鉴别诊断奠定了基础。  相似文献   

H7N2禽流感病毒分离株血凝素蛋白全基因的序列分析     

王传彬 孙明田克恭  金萍王宏伟  遇秀玲陈西钊 《中国兽医科技》2005,35(12):988-993

为进一步分析H7N2禽流感病毒（AIV）分离株血凝素（HA）基因的特性,参照已发表序列设计了1对引物,采用RT-PCR获得了1条约1.7 kb的DNA片段,测序后进行了同源性比较、HA基因系统发育进化树分析和氨基酸编码分析.结果表明,所测的2个分离株的HA基因全长1 664 bp,编码除信号肽以外的HA蛋白的全部544个氨基酸,其中包括HA1的323个氨基酸,HA2的221个氨基酸.2个分离株HA基因核苷酸序列的同源性为99.4%;与GenBank中AIV标准株A/Afri.Star./Eng-Q/983/79（H7N1）的同源性最高,分别为99.4%和99.0%;与美国A/Chicken/NewYork/13142-5/94（H7N2）株同源性很低（仅65.0%）,而与以色列、意大利H7N2 AIV的同源性较高（为96%～97%）;2个分离株在HA基因进化树中均处于H7亚型AIV的欧亚群系分支内.推导氨基酸的序列分析表明,其HA蛋白裂解位点的氨基酸序列为-GR-GLF-,仅包含1个碱性氨基酸（R-）残基,符合低致病力AIV的基因特征.  相似文献   

Comparison of morphology, viability, and function between blood and milk neutrophils from peak lactating goats          下载免费PDF全文

Sui Zhi Tian  Chai Ju Chang  Chih Chi Chiang  Huo Cheng Peh  Mu Chiou Huang  Jai-Wei Lee    Xin Zhao 《Canadian journal of veterinary research》2005,69(1):39-45

The morphological features of blood and milk neutrophils from peak lactating goats were compared using light microscopy, scanning electron microscopy and flow cytometry in order to investigate the cytological changes of neutrophils after migration into the mammary gland. The kinetics of reactive oxygen intermediates (ROI) generation and gelatinase release of blood and milk neutrophils, with or without stimulation of phorbol 12-myristate, 13-acetate ester (PMA), were used to characterize their responses to inflammatory stimuli. Neutrophils isolated from goat milk were highly segmented and contained multi-lobed nuclei. Ultrastructurally, milk neutrophils were more ruffled on the surface compared to blood neutrophils. Approximately 30% of milk neutrophils were undergoing cell death, either necrosis or apoptosis, in contrast to 8% of blood neutrophils. The ROI production of activated milk neutrophils peaked earlier than blood neutrophils, but the duration and the intensity were much less. Neutrophils from both sources augmented the release of gelatinase in response to PMA (1 ng/mL). However, the amount of gelatinase released from milk neutrophils was lower (P < 0.05) than that of blood neutrophils. In summary, more neutrophils become apoptotic and necrotic in the mammary gland, presumably due to spontaneous aging, the process of diapedesis, and the interaction with milk components. Milk neutrophils have impaired functionalities in comparison with blood neutrophils. The information is relevant when studying mammary gland immunity and related diseases, such as mastitis.  相似文献   

High-throughput real-time RT-PCR assay to detect the exotic Newcastle Disease Virus during the California 2002--2003 outbreak.   总被引：4，自引：0，他引：4  

Beate M Crossley  Sharon K Hietala  Liu-Mei Shih  Lou Lee  Evan W Skowronski  Alex A Ardans 《Journal of veterinary diagnostic investigation》2005,17(2):124-132

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Culture of Differentiated Adult Rabbit Auricular Chondrocytes     

V. Villar  B. Colao  I. Calles‐Venal  J. G. Fernndez‐lvarez  M. Fernndez‐Caso  J. M. Villar 《Anatomia, histologia, embryologia》2005,34(Z1):54-54

Chondrocytes dedifferentiate to a fibroblast‐like phenotype on plastic surfaces. Dedifferentiation is reversible if these cells are then cultured embedded in gels as alginate, agarose or collagen. Chondrocytes cultured in suspension on a non‐adherent surface are also known to form aggregates of differentiated cells. The knowledge of chondrocyte behavior in culture is relevant for tissue engineering purposes. In this report we describe a simple method to culture differentiated or redifferentiated rabbit auricular chondrocytes on plastic surfaces with a stable phenotype. When chondrocyte aggregates formed in suspension are next seeded on plastic surfaces, most of them attach to the plastic as round or polygonal cells, and this morphological differentiation, confirmed by the presence of type II collagen, is stable for long culture periods. We also report that the addition of aggregates to monolayer cultures of dedifferentiated chondrocytes results in their redifferentiation, as is shown by their morphological changes and the synthesis of type II collagen. Therefore, this simple method can be useful for the study of chondrocyte behavior on plastic surfaces and for redifferentiating previously proliferated chondrocytes in tissue engineering techniques. Furthermore, these results demonstrate that, in addition to culture conditions such as cell isolation method or cell‐density, chondrocyte behavior on plastic depends on the presence or absence of aggregates resulting from the dissociation process.  相似文献   

韩国一次性收获的红椒品种—Saeng-ryeog216     

M.C.Cho  D.H.pae  Y.S.Cho  Y.Chae  W.M.Lee  D.H.Kim 《辣椒杂志》2003,(4)

根据栽培面积以及为农民提供的收入而言,红辣椒在韩国一直是仅次于水稻而居第二位的重要农作物。按人均计算,韩国人均年辣椒粉的消费量达2.5kg以上,可能是世界最高记录。辣椒在韩国主要用作泡菜(韩国称之为Kimchi)香辛料和菜肴中的  相似文献   

不同地下水位植物蒸腾耗水特性研究   总被引：20，自引：6，他引：20  

赵明  郭志中  王耀琳  李爱德  张德魁  贾宝全 《干旱区研究》2003,20(4):286-291

在甘肃省民勤治沙站利用非称量测渗仪研究了梭梭、柠条等 10种沙区植物的蒸腾耗水特征。结果表明 ,各种植物 5a的总蒸腾量由大到小排列为 :沙枣、花棒、沙木蓼、柠条、梭梭、白皮沙拐枣 ;随植物年龄增长 ,蒸腾系数有上升的趋势 ,而蒸散系数有下降的趋势 ;在民勤沙区 6～ 9月是主要蒸腾季节 ,该时段的蒸腾量占全年蒸腾量的80 %～ 90 % ;植物的蒸腾量随年龄的增长而迅速增加 ,但增长幅度随物种不同而异。一般生长快的植物增长快 ,生长慢的植物增长慢。  相似文献   

‘三棱榄''橄榄果实香气成分分析   总被引：7，自引：1，他引：7  

钟明  陈玉芬  甘廉生  蔡时可 《园艺学报》2003,30(6):757-761

1 材料与方法选取广东优良鲜食橄榄品种‘三棱榄’,2001年12月6日采样,采用固相微萃取法(SPME)富集香气成分(鲜橄榄果肉于15℃下捣碎后取样1.0 g放入4 mL聚四氟乙烯硅橡胶垫密封螺口玻璃瓶中,插入100μm聚二甲基硅氧烷纤维头于室温25-30℃顶空取样2 h),用美国Finnigan TRACE GC-MS气相色谱-质谱联用仪进行分析。气相色谱柱为DB-1弹性毛细管柱30 m×0.25 mm,载气为He(99.99％),流速1.0mL/min。程序升温从40℃开始先保持10 min,后以2℃/min的升温速率升至150℃保持10 min。质谱条件:电子能量70 eV,离子源温度250℃,质量范围35-450 aum,不分流进样。2002年12月18日采样重复分析。  相似文献   

Annual bluegrass (Poa annua L.) control in bentgrass (Agrostis palustris Huds.) green with sequential application of bispyribac‐sodium combined with dinitroanilines*     

NAM‐IL PARK  YUKO SUTO  YUTAKA MIURA  NOBUO NAKATANI  SHIN‐ICHI IORI  MASARU OGASAWARA 《Weed Biology and Management》2002,2(3):159-162

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巴旦杏在甘肃河西地区引种培育问题的探讨     

牛云  赵明  张宏斌  赵国生  雷军  胡跃军 《北方果树》2002,(6):22-24

该文通过分析巴旦杏在甘肃河西地区引种培育过程中出现的问题,总结出用容器育苗和大田开沟点播相结合来培育接穗,以山桃为砧木嫁接巴旦杏的引种驯化技术.结果表明,容器培育接穗比直接大田点播出苗率、成活率分别提高了10%和15%;巴旦杏与山桃嫁接表现出极好的亲和性,嫁接成活率为90%.  相似文献