Detection,cloning and expression of bone morphogenetic protein-1 from human osteosarcoma cell lines     

CHEN Dong-ying  ZHU Quan-sheng  LIU Chao  QIU Ju-shi 《园艺学报》2003,19(6):751-756

AIM:To manufacture recombinant protein of the highly conserved domain in human bone morphogenetic protein-1(BMP-1) using gene engineering methods as antigen for making wide spectrum antibody to BMP-1.METHODS:We analyzed the gene sequences and protein structures of BMP-1 and its related proteins, and chose a highly conserved fragment as target gene. Total RNA was prepared from human osteosarcoma cell line Saos-2, then the target gene was amplified with RT-PCR. The PCR product was cloned into prokaryotic expression vector pMAL c2 to get recombinant vector BMP-1(322-588aa)-pMAL c2. After transforming the recombinant plasmid into DH5-alpha and screening, several prositive clones were got for sequencing. Finally the transformed cells was induced with IPTG to get fusion protein.RESULTS:The BMP-1 gene fragment was successfully cloned into vector pMAL c2, and was able to express efficiently with IPTG inducement. The amount of expressed fusion protein is about 66%-72% in total volume of bacterial proteins.CONCLUSIONS:The recombinant protein contains several key domains(2 CUB domains and 1 EGF domain), which are shared by BMP-1 and its related proteins. Specific wide spectrum antibody to human BMP-1 and its related proteins may be generated with this recombinant protein antigen.  相似文献   

Effect of hemin on the expression of heme oxygenase-1 in rat vascular smooth muscle cells     

HU Chang-xing  MA Ye-xin  FU Du-guan  REN Peng 《园艺学报》2003,19(11):1493-1496

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Engineering of a Single Chain Variable Fragment Antibody Specific for the Citrus tristeza virus and its Expression in Escherichia coli and Nicotiana tabacum     

P. Galeffi  G. Giunta  S. Guida  C. Cantale 《European journal of plant pathology / European Foundation for Plant Pathology》2002,108(5):479-483

Citrus tristeza virus (CTV) is one of the most destructive citrus virus diseases in the world. The construction of an engineered antibody, EMBL accession number AJ278109, able to specifically recognize its antigen, i.e. the coat protein of CTV, directly on infected plant material without any purification or manipulation of the entire woody plant. The potential uses of this engineered antibody are discussed.  相似文献   

Citrus variegation virus: Molecular Variability of a Portion of the RNA 3 Containing the Coat Protein Gene and Design of Primers for RT-PCR Detection     

Bahia Bennani  Celso Mendes  Mustapha Zemzami  Houssine Azeddoug  Gustavo Nolasco 《European journal of plant pathology / European Foundation for Plant Pathology》2002,108(2):155-162

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Detection of Colletotrichum coccodes from soil and potato tubers by conventional and quantitative real-time PCR   总被引：4，自引：1，他引：4  

D. W. Cullen  A. K. Lees  I. K. Toth  J. M. Duncan † 《Plant pathology》2002,51(3):281-292

Colletotrichum coccodes is the causal agent of the potato blemish disease black dot. Two PCR primer sets were designed to sequences of the ribosomal internal transcribed spacer (ITS1 and ITS2) regions for use in a nested PCR. The genus-specific outer primers (Cc1F1/Cc2R1) were designed to regions common to Colletotrichum spp., and the species-specific nested primers (Cc1NF1/Cc2NR1) were designed to sequences unique to C . coccodes . The primer sets amplified single products of 447 bp (Cc1F1/Cc2R1) and 349 bp (Cc1NF1/Cc2NR1) with DNA extracted from 33 European and North American isolates of C. coccodes. The specificity of primers Cc1NF1/Cc2NR1 was confirmed by the absence of amplified product with DNA of other species representing the six phylogenetic groups of the genus Colletotrichum and 46 other eukaryotic and prokaryotic plant pathogenic species. A rapid procedure for the direct extraction of DNA from soil and potato tubers was used to verify the PCR assay for detecting C. coccodes in environmental samples. The limit of sensitivity of PCR for the specific detection of C. coccodes when inoculum was added to soils was 3·0 spores per g, or the equivalent of 0·06 microsclerotia per g soil, the lowest level of inoculum tested. Colletotrichum coccodes was also detected by PCR in naturally infested soil and from both potato peel and peel extract from infected and apparently healthy tubers. Specific primers and a TaqMan fluorogenic probe were designed to perform quantitative real-time (TaqMan) PCR to obtain the same levels of sensitivity for detection of C. coccodes in soil and tubers during a first-round PCR as with conventional nested PCR and gel electrophoresis. This rapid and quantitative PCR diagnostic assay allows an accurate estimation of tuber and soil contamination by C. coccodes .  相似文献   

干旱区自然灾害监测预警系统的一般模式——以塔里木盆地为例     

王让会  卢新民 《干旱区资源与环境》2002,16(4):64-68

我国西部干旱区生态环境脆弱 ,自然灾害频繁。塔里木盆地是我国最大的内陆盆地 ,干旱、大风、沙尘暴、洪水、泥石流以及地震、雪灾等自然灾害 ,严重地影响到盆地的生态安全。在RS、GIS及 GPS等技术支持下 ,研究各自然灾害的孕灾机理及过程 ,建立自然灾害的监测评价及预警系统 ,对于塔里木盆地生态环境建设具有重要意义。  相似文献   

Bt对小地考虑呼吸作用的影响   总被引：5，自引：0，他引：5  

李芳  谌建春等 《昆虫天敌》2002,24(1):15-19

利用红外CO2分析仪测定小地老虎幼虫呼吸节律的变化结果显示：小地老虎幼虫呼吸节律属于不规则波动型，取食含Bt烟叶的幼虫呼出的CO2量显著低于对照组，且呼吸节律的波动性也明显增大，说明小地考虑幼虫取食Bt后，呼吸代谢受到一定干扰，出现呼吸水平降低及代谢紊乱现象。  相似文献   

ITS—RFLP分析进行昆虫病原线虫斯氏属和异小杆属的分类鉴定   总被引：2，自引：1，他引：2  

李菁菁  韩日畴等 《昆虫天敌》2002,24(3):97-104

本研究通过PCR扩增和六种限制性内切酶（AluⅠ，HinfⅠ，MboⅠ，RsaⅠ，HaeⅢ和PvuⅡ）酶切，对国内害虫防治上常用的几种昆虫病原线虫，包括斯氏属S.car-pocapsae,S.feltiae和S.glaseri以及异小杆属H.bacteriophora,H.zealandica,H.indicus和H.megidis等8个品系rDNA-ITS进行分析。建立起可以区分各线虫种的标准RFLP图谱。该方法快速简便，稳定可靠，需要的样品量少。可以用于新鲜的，或冻存的样品，甚至分析单条的线虫，不仅可进行昆虫病原线虫的快速分类鉴定。而且进一步可以应用于线虫田间释放的辅助监测。实际田间感染率的测定和线虫毒力的比较。  相似文献   

新型取代氨基(或芳氧)磺酰基苯氧丙酸酯的合成及其除草活性   总被引：3，自引：0，他引：3  

胡方中  吕凯  任雪玲  陈彬 《农药学学报》2002,4(4):17-22

由α-苯氧基丙酸酯出发,合成了取代氨基(或芳氧)磺酰基苯氧丙酸酯,并测定了它们的除草活性。所有的化合物均经1H NMR和元素分析确证,初步的生测结果表明上述化合物具有一定的除草活性。  相似文献   

麦蚜对吡虫啉的敏感性及吡虫啉有效用量的评价   总被引：1，自引：0，他引：1  

刘爱芝  王晓军  李素娟  武予清  李巧丝 《植物保护》2002,28(3):15-18

室内测定结果表明 ,禾谷缢管蚜对吡虫啉的敏感性比麦长管蚜高 ,其LC₅₀之比为1∶3左右。田间试验结果明确 ,吡虫啉持效性好 ,其持效期与吡虫啉用量和麦蚜种类有关。小麦穗蚜发生始盛期是吡虫啉防治麦蚜的最佳时期 ,防治禾谷缢管蚜最佳有效用药量为1g/667m²,麦长管蚜为2g/667m²。  相似文献