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   

转基因苜蓿研究进展   总被引：4，自引：0，他引：4  

胡清泉  玉永雄 《中国草地》2005,27(5):58-62

就苜蓿基因工程在育种上的研究、转基因技术在苜蓿上的进展做了简要介绍。  相似文献   

山区农业面源污染的防治途径及对策     

喻尚其刘光德  李其林赵中金 《四川畜牧兽医学院学报》2005,3(4):1-4

结合重庆市地形地貌特点和山区农业面源污染发生的因子，提出山区农业面源污染的防治对策。一方面必需结合当地的自然环境条件和经济发展水平提出农业面源污染的防治规划；另一方面必需结合生态农业建设这一主线，使物质、能量在生态系统内良性循环。紧紧抓住产生农业面源污染源的因子——化肥、农药、农田秸杆、畜禽粪便、农村生活废物等，提出科学的防治和减少途径。  相似文献   

吉林省荒地资源生态适宜性评价     

张妍  尚金城  于相毅 《干旱区资源与环境》2003,17(3):35-38

本文从吉林省荒地资源的实际出发 ,在荒地资源生态适宜性评价基本理论的指导下 ,采用地理信息系统方法 ,对吉林省荒地资源进行了生态适宜性评价。通过空间数据和属性数据的融合、分析、处理 ,可以直观获得荒地资源的生态适宜度等级 ,从而提高评价效率 ,为荒地资源的合理开发利用提供科学依据。  相似文献   

乌梁素海湿地环境与资源地理信息系统的研究   总被引：6，自引：0，他引：6  

尚士友  吴利斌  郁志宏  岳海军  李洁  马清艳 《干旱区资源与环境》2003,17(5):59-64

本系统建立乌梁素海湿地基础地形库、生态库、资源库、工程档案库、遥感影像库等基础数据库及信息发布系统 ,使 GPS、GIS、RS紧密结合 ,同时实现了专家决策支持系统与地理信息系统之间的一体化集成。本系统可以为乌梁素海湿地生态治理与资源开发提供全方位的数据信息及其管理 ,对生态工程规划与实施方案进行优化设计 ,并提供实时监控服务。  相似文献   

Increased expression of CD40 ligand by peripheral blood mononuclear cells in patients with systemic lupus erythematosus     

LI Yu-jie  LI You-ji  YE Ren-gao  KONG Qing-yu  DONG Xiu-qing  YU Xue-qing 《园艺学报》2003,19(4):514-517

AIM:To investigate expression and function of CD40 ligand by peripheral blood mononuclear cells (PBMCs) in patients with systemic lupus erythematosus (SLE).METHODS:Expression of CD40 ligand by PBMCs in patients with SLE and control were examined by flow cytometric analysis before and after stimulated by phytohemagglutinin(PHA)and depressed by Dexamethasone(Dex). The correlation between expression of CD40 ligand and SLE activity index(SLEDAI) was analysed in patients with SLE.RESULTS:The expression of CD40 ligand by PBMCs in patients with active SLE was higher than that in patients with inactive SLE and control. Though the expression of CD40 ligang by PBMCs could be stimulated by PHA in three groups, it was the highest in patients with active SLE. Dex depressed the expression of CD40 ligand by PBMCs significantly in patients with SLE, but not in control. There was high positive correlation between expression of CD40 ligand and SLEDAI in patients with active and inactive SLE.CONCLUSION:Increased expression of CD40L by PBMCs in patients with SLE may play an important role in pathogenesis of SLE.  相似文献   

Association of polymorphism of Fc receptor γ chain gene at position-29 in promoter with the susceptibility to systemic lupus erythematosus in southern Chinese     

ZHOU Shu-lu  YE Ren-gao  LIU Xiao-bo  ZHANG Hong  XU Han-shi  DU Yong  LI You-ji  YANG Nian-sheng  YANG Xiao  YU Xue-qing 《园艺学报》2003,19(6):782-785

AIM:To detect the association between the polymorphism of Fc receptor γ chain gene at position-29 in promoter and systemic lupus erythematosus(SLE).METHODS:The genotypes at position -29 in promoter of Fc receptor γ chain gene were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method in 180 patients with SLE and 140 ethnically matched controls in southern China.RESULTS:The frequencies of TT genotype(33.3%) and T allele (54.4%) at position -29 in patients with SLE were significantly higher than those in controls (17.2% and 42.9%, respectively), whereas, the frequencies of GG genotype (24.4%) and G allele (45.6%) in patients with SLE were remarkably lower than those in controls (31.4% and 57.1%, respectively) (P＜0.05). The TT genotype and T allele at position -29 were not associated with lupus nephritis in SLE patients (P＞0.05).CONCLUSION:Our results indicate that the T allele at position -29 in promoter of Fc receptor gene probably contributes to the susceptibility to SLE, but does not play a role in the occurrence of lupus nephritis.  相似文献   

Using antisense nucleic acid technology to study the influence of POLκ on genetic stability     

LUO Yue-qiu  YU Ying-nian 《园艺学报》2003,19(9):1153-1156

AIM:To establish cell line FL-POLκ^- and to study the role ofPOLκ(polymerase kappa) on genetic stability.METHODS:A mammalian expression vector expressing antisense POLκ gene fragment pMAMneo -amp^--POLκ was constructed by cloning the 1 690-1 918 fragment of POLκ gene into the mammalian expression vector pMAMneo-amp^- in antisense orientation. FL cells were fransfected with this antisense RNA expressing vector and selected by G418. Based on the shuttle-plasmid pZ189, the mutation assay was made.RESULTS:The spontaneous mutation frequency of supF tRNA gene in the plasmid replicated in the FL- POLκ^- was 11.2×10^-4, while it was 4.9×10^-4 and 3.7×10^-4 in the control cells FL and FL-M, respectively.CONCLUSION: POLκ playes an important role in maintenance of genetic stability.  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