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马明

作品数:14 被引量:70H指数:5
供职机构:北京市农林科学院畜牧兽医研究所更多>>
发文基金:北京市农林科学院青年基金北京市优秀人才培养资助北京市自然科学基金更多>>
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14 条 记 录,以下是 1-10
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鸡、鸽、虎呼吸道和消化道黏膜上皮细胞表面流感病毒受体类型的检测被引量:6
2009年
【目的】检测鸡、鸽和虎体内的流感病毒受体的类型和分布,以期解释3种动物对禽流感病毒易感性差异的机制。【方法】使用地高辛标记的外源性凝集素染色方法检测这些动物的喉头、气管、肺脏和肠道(直肠)的上皮细胞表面SAα2,6Gal和SAα2,3Gal连接键的类型。【结果】SPF鸡的上呼吸道黏膜上皮细胞表面有大量的SAα2,3Gal和少量的SAα2,6Gal;肺房上皮细胞表面只有SAα2,3Gal;而直肠黏膜上皮中SAα2,6Gal和SAα2,3Gal都没有表达。成年鸽的上呼吸道黏膜上皮细胞表面只有SAα2,6Gal,而没有SAα2,3Gal;而下呼吸道中SAα2,6Gal和SAα2,3Gal都没有;直肠黏膜上皮细胞只有SAα2,3Gal。虎的呼吸道和消化道(直肠)黏膜上皮细胞表面有大量的SAα2,6Gal和SAα2,3Gal。【结论】鸡和虎具有禽流感病毒受体,对禽流感病毒易感,鸽不具备禽流感病毒受体,因此,鸽对禽流感病毒不易感。
马明刘月焕陈明勇韩春华林健
关键词:唾液酸
传染性法氏囊病病毒VP5基因的克隆与原核表达被引量:3
2008年
根据GenBank已登录的传染性法氏囊病病毒(IBDV)VP5基因序列,设计合成了一对VP5基因特异性引物,应用RT-PCR技术从IBDV标准毒株中扩增得到VP5基因,并将其克隆到原核表达载体pGEX-6P-1中,构建了重组原核表达质粒pGEX-6P-1-VP5,对重组表达质粒鉴定正确后,转化大肠埃希菌BL21进行诱导表达,SDS-PAGE和Western blot检测结果表明,IBDV VP5基因在大肠埃希菌BL21中得到了正确表达,所表达的融合蛋白与IBDV阳性血清具有特异性抗原抗体反应。
吴琼马明何桂梅陈明勇
关键词:传染性法氏囊病病毒VP5基因基因克隆
Protective Efficacy of Newcastle Disease Vaccines against Prevalent Strains in Recent Years
2009年
[Objective] To investigate the protective efficacy of currently available Newcastle disease vaccines against currently prevalent strains and thus provide guidance for the application of vaccines and the prevention and control of Newcastle disease. E Method] The 6-week-old SPF chickens were respectively inoculated with five live attenuated Newcastle disease vaccines (A, B, C, D, and E) at the dose of 1-fold or 0.01-fold usage via intranasal, intraocular or oral route. After 14 d post immunization, the titers of HI antibody were detected. And all the chickens were chal- lenged by Newcastle disease virus (NDV) FEo standard strain or the isolated wild strains, NDV-2007-HB and NDV-2008-YB. The clinical symptoms of chickens were continuously observed, and the morbidity and mortality were determined. [ Resalt] After 14 d post immunization, antibodies were induced at a protective level in chickens immunized with the five vaccines. As shown by the animal experiment, the five vaccines at the dose of 1-fold or 0.01-fold usage protected all vaccinated chickens from the death caused by NDV strain, and more than 90% of vaccinated chick- ens from the death caused by NDV-2007-HB strain, while the vaccines, A, B, and C, at the dose of 0.01 -fold usage protected more than 90% of vaccinated chickens from the death caused by NDV-2008-YB strain. E Conclusion] Under laboratory conditions, the currently available Newcastle disease vaccines have better protective efficacy against the two currently prevalent NDV strains and prevent the occurrence of Newcastle disease.
林健刘月焕韩春华马明刘永宏潘洁
鸽等动物禽流感流行病学与免疫防治的研究
刘月焕韩春华林健祝俊杰赵德明宋维平李秀敏步卫东姚炜光杨兵姜北宇韦海涛郑瑞峰孟颜妮马明
该项目属农业(兽医)科学技术领域。该项目属于社会公益类研究项目,主要研究内容为(1)北京市野生动物、猪、珍禽等动物高致病性禽流感的流行病学调查;(2)鸽子对H5N1亚型禽流感病毒易感性的研究;(3)鸽子对H5N1亚型禽流...
关键词:
关键词:免疫防治养鸽业
Location of Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus in Tissues of Natural Cases被引量:15
2009年
[ Objective] The aim of this study was to provide a theoretical basis for the prevention and treatment of highly pathogenic porcine reproductive and respiratory syndrome (HP-PRRS). [Method] Antigen location and histopathological observation in natural cases infected by highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) were analyzed by immunohistochemistry and H. E. staining. [Result] The virus antigen mainly existed in epithelial calls, and also a few in mecrophages, lymphocytes and brain nerve cells. [ Conclusion] The cell and tissue tropism of HP-PRRSV strain in natural cases is different from that of previous strains.
刘永宏赵丽韩春华王凤龙刘月焕林健杨汉春郭鑫李栋梁韦海涛祝俊杰赵景义赵振华马明杨龙峰王金玲
关键词:IMMUNOHISTOCHEMISTRY
Sequence Analysis of HA Genes from Three H9N2 Subtype Avian Influenza Viruses被引量:2
2009年
[ Objective] The study aimed to understand the genetic characters of H9N2 subtype avian influenza viruses isolated in Belling area. [ Method] HA genes of three H9N2 subtype avian influenza viruses A/Chicken/Beijing/xu/00, A/Chicken/Beijing/bei/00 and A/Chicken/Beijing/ liu/00 were amplified by RT-PCR and then sequenced. [ Result] The results of phylogenetic analysis showed that A/Chicken/Beijing/xu/00, A/ Chicken/Beijing/bei/00 and A/Chicken/Beijing/liu/00 shared the nucleotide homologies of 84.8% ( Dk/HK/Y439/97 ) -98.0% ( Ck/GX17/00 ), 85.1% (Dk/HK/Y439/97) - 99.1% ( Ck/GXl 7/00), 90.7% ( Ck/BJ/3/01 ) - 99.1% (Ck/GX17/00) with the isolates from Hongkong and other are- as of Chinese Mainland respectively. At the same time, the analysis of amino acid indicated that the three isolates belonged to low pathogenic H9N2 isolates of avian origin. The 226^th amino acid of them were L ( Leu), suggesting their high binding affinity to human cells. There were seven glyco- sylation sites in HA protein, five from HA1 and two from HA2. [ Cenclusien] By analysis at molecular level, it could be concluded that A/Chicken/ Beijing/xu/00, A/Chicken/Beijing/bei/00 and A/Chicken/Beijing/liu/00 were low pathogenic H9N2 isolates of avian origin.
韩春华林健刘月焕潘洁马明刘永宏
虎呼吸道和消化道上皮细胞表面流感病毒受体种类的测定
流感病毒颗粒表面的红细胞凝集素(HA)经与宿主细胞膜上多糖末端的唾液酸结合后附着于细胞表面,随后细胞膜内陷并逐步包裹病毒颗粒,通过胞饮作用将病毒吞入,完成病毒感染细胞的第一步。流感病毒唾液酸受体连接键型有两种,分别是唾液...
马明刘月焕陈明勇韩春华林健
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Expression of Porcine Reproductive and Respiratory Syndrome Virus ORF7 Gene and Purification and Immunological Activity Analysis of the Recombinant Protein被引量:14
2009年
[Objective] The aim of this study was to realize efficient expression of the porcine reproductive and respiratory syndrome virus (PRRSV) ORF7 gene in genetic engineering bacteria and analYze the immunological activity of the recombinant protein after purification. [ Method] The constructed recombinant expression vector pET-ORF7 was transformed into Escherichia co1BL21 (DE3) and induced by IPTG under the optimal condition. After analysis of SDS-PAGE and Western Blot, the expression products were purified by Ni-NTA His · Bind Resin chrom- atographic column under denaturing condition and renatured by gradient dialysis. Subsequently, the immunological activity of the renatured recombinant protein was detected by Westem Blot and indirect ELISA. [ Result] The recombinant plasmid pET-ORF7 expressed in E. coli successfully, and the fusion protein was in the form of inclusion body. By SDS-PAGE detection, the molecular weight of the expression protein was approximate 33 kD, according with the expectation. Analysis by Bandscan software showed that the expressed fusion protein was about 50% of total bacterial protein of BL21 (DE3). Wastem Blot and indirect ELISA detection showed that the renatured protein could react with PRRSV positive serum specifically, indicating its good immunological activity. [ Conclusion] This study lays a foundation for the preparation of PRRSV monoclonal antibody and diagnostic kit.
张永富韩春华林健刘月焕韦海涛祝俊杰赵景义李栋梁马国文布日额李明刚张婷刘永宏马明张秋雨
关键词:EXPRESSIONPURIFICATION
北京3株H9N2亚型禽流感病毒HA基因的序列分析被引量:6
2009年
[目的]为了解2000年北京地区H9N2分离毒株的遗传特点。[方法]采用RT-PCR技术扩增了3株H9N2亚型禽流感病毒北京地区分离株的HA基因片段,并对所得序列进行分析比较。[结果]遗传演化分析表明,所分离的3个毒株的HA基因与其他中国内地和香港毒株的同源性分别为XU株84.8%(Dk/HK/Y439/97)~98%(Ck/GX17/00),LIU株85.1%(Dk/HK/Y439/97)~99.1%(Ck/GX17/00),BEI株90.7%(Ck/BJ/3/01)~99.1%(Ck/GX17/00)。氨基酸序列分析发现,3株病毒均为禽源低致病力毒株,226位氨基酸均为L(Leu),更易于与人类细胞结合;HA蛋白存在7个糖基化位点。[结论]从分子水平分析,A/Chicken/Beijing/xu/00、A/Chicken/Beijing/bei/00和A/Chicken/Beijing/liu/00株属于低致病力的H9N2亚型禽源毒株。
韩春华林健刘月焕潘洁马明刘永宏
关键词:H9N2亚型禽流感病毒HA基因
新甲型H1N1流感病毒的分子遗传进化分析被引量:3
2010年
为了分析2009年新型A型H1N1流感病毒分子流行病学的特点,本文从NCBI上下载不同分离宿主的流感毒株各节段全基因序列应用分子生物学软件进行遗传演化分析,结果显示:新型A(H1N1)流感毒株与过去的人源A(H1N1)毒株相比差异较大,相对于此前的人源A(H1N1)流感毒株,HA出现了79个位点发生突变。其中14个是相对于所有来源A(H1N1)流感毒株的新突变位点,但有37个突变位点只能在猪源毒株中找到。究竟这一现象说明新型A(H1N1)流感毒株是猪、人源毒株的重组变异毒株,还是猪源H1N1感染人群后逐渐变异并适应人群的结果,还有待进一步研究。
赵丽刘永宏刘月焕王凤龙林健韩春华马明丁玉林丁旭娜王金玲杨龙峰潘洁韩婧雯
关键词:甲型流感遗传演化分析
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