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1.
转基因玉米特异性检测阳性标准分子的构建与应用   总被引:3,自引:1,他引:2  
试验用PCR方法从玉米中扩增内源基因zSSIIb片段,并将其克隆到pMD18-T载体上,获得中间载体pMD-zSSIIb;根据Bt11和MON810玉米转化体特异性序列,分别设计带酶切位点的引物,扩增出Bt11和MON810转化体特异性产物;用相应的酶对pMD-zSSIIb和两种PCR产物进行酶切,分别将Bt11和MON810产物克隆到pMD-zSSIIb上,获得阳性标准分子pMD-ZB和pMD-ZM,并对其进行特异性测试。结果表明,获得的阳性标准分子可以作为转基因产品检测时的阳性对照。  相似文献   

2.
以若干定性PCR方法部颁标准对含有0.5%的4份不同转基因混合样品进行检测,先以通用元件标准中的CaMV35s启动子、NOS终止子对混合样品进行初步定性PCR筛选。结果表明,4份样品中都含有转基因成分。Bt基因特异性标准检测表明,3#和4#样品含有转基因抗虫水稻成分。构建特异性标准PCR检测表明,2#、3#和4#样品含有转基因GTS-40-3-2大豆成分。以MON810、Bt176、NK603转化体事件标准进行品系特异性PCR检测,结果证实:1#和4#样品中含有Bt176转基因玉米成分;3#样品中含有Mon810转基因玉米成分;4份样品中均不含NK603转基因玉米成分。说明农业部颁布的定性PCR方法标准能满足于对多种转基因混合样品的检测,且检测结果准确,可靠。  相似文献   

3.
转基因玉米MON810(Yield Gard R)是孟山都公司通过DNA重组技术和微注射轰击研发的一种具有对欧洲玉米螟(ECB;Ostrinia nublialis)有特殊抗性的转基因玉米品系,目前在世界各地已经得到广泛种植,为加强对该品系玉米的安全管理,本研究旨在建立MON810品系玉米的转化事件特异性定性PCR检测方法。根据MON810的插入序列信息,在3′端的侧翼序列处设计定性PCR检测的引物,检测MON810在其他几种常见转基因作物混合样品的特异性。结果表明,该方法具有很好的特异性。同时检测该引物系统的扩增灵敏度,结果表明,检测引物的灵敏度可达0.1%。建立的MON810特异定性PCR检测方法经全国7家实验室的验证,进一步证实该方法能够特异地检测出样品中的MON810转化事件,检测方法的灵敏度可达0.1%,且检测结果具有良好的可重复性和可重现性。MON810转化事件定性PCR检测方法的建立可满足于抗虫转基因玉米MON810及其衍生品种生物安全管理的需要。  相似文献   

4.
抗虫玉米MON89034转化体特异性PCR检测技术研究   总被引:5,自引:2,他引:3  
根据抗虫玉米MON89034外源插入片段5’端与植物基因组连接区序列设计特异性引物,并进行PCR扩增,预期产物大小为455 bp。以zSSIIb基因作为内标准基因,建立了转基因玉米MON89034转化体特异性定性PCR检测方法。对该方法进行重现性、特异性和灵敏度测试,结果表明:该方法能够特异性检测出MON89034转化体,以100 ngDNA为模板,该方法的检测灵敏度达到0.1%,约为40个起始模板拷贝。复合PCR检测结果还表明,在同一PCR反应管中可实现对zSSIIb基因和MON89034的同时检测。  相似文献   

5.
采用PCR技术检测CaMV 35S启动子,并进一步通过PCR检测RoundUp Ready Soybean(RRS)和Btl76 Maximaizer的特异性DNA片段,判断大豆和玉米加工产品中是否含相应转基因成分.在1份豆粕和豆腐样品中检测到了RoundUp Ready大豆特异性的498 bp片段,而在玉米粒样品中检测到了Btl76特异性转基因成分.PCR检测的灵敏度达到0.1%,稳定性良好.结果表明,PCR技术检测外源基因是灵敏和准确的,可以广泛地应用到转基因作物及其加工产品的转基因成分检测中.  相似文献   

6.
应用单管巢式和半巢式PCR检测转基因玉米MON89034   总被引:1,自引:1,他引:0  
根据MON89034玉米的5’端和3’端边界序列分别设计1组转化体特异性的巢式PCR引物,采用中途进退式PCR策略建立MON89034玉米的转化体特异性检测方法,扩增产物分别为491 bp和188 bp。以转基因玉米MON89034及8种其他转基因作物为材料,证明此方法对MON89034玉米具有高度特异性。灵敏度测试结果表明,此方法的相对检出限达到0.01%,绝对检出限为4个单倍体基因组拷贝数,比普通PCR提高了5倍。建立的单管巢式和半巢式PCR方法可准确、高效地检测转基因玉米MON89034及其产品。  相似文献   

7.
本文参考出入境检验检疫行业标准中转基因油菜外源基因的PCR检测体系,分析了18种不同DNA聚合酶对定性PCR检测转基因油菜OXY235中的CaMV35S启动子的影响。结果表明,不同DNA聚合酶对CaMV35S启动子定性PCR检测中非特异扩增的产生以及检测灵敏度影响较大。其中热启动Taq酶效果较好,目标条带清晰,检测灵敏度高,而且完全没有非特异性扩增条带,非常适合用于定性PCR检测。对于同样条件下无法有效扩增0.1%DNA样品的Taq酶,则不推荐用于检测途径。  相似文献   

8.
应用聚合酶链式反应技术(PCR)扩增了玉米淀粉分支酶的cDNA基因片段,并将其克隆到pMD18-Tvector载体上,对重组子进行PCR检测和限制性内切酶分析,并测定了DNA全序列.结果表明:克隆片段全长为1 698 bp.将该基因反向插入到pCAMBIA1301的CaMV35S启动子之后,构建了反义表达载体.  相似文献   

9.
环状等温扩增技术快速检测转基因玉米MON863的研究   总被引:3,自引:1,他引:2  
兰青阔  王永  赵新  朱珠  程奕 《玉米科学》2011,19(1):31-34
采用环状等温扩增技术(Loop-Mediated Isothermal Amplification,LAMP),针对转基因玉米品系MON863中玉米基因组与外源基因结合处设计4条特异性引物,建立MON863的快速检测技术体系,并对时间、温度等反应条件及体系灵敏度、特异性等进行探索。结果表明:该检测体系最适反应温度为63℃,反应时间60 min,灵敏度是常规定性PCR的10倍。该检测方法具有高度的特异性和稳定性,操作方便、简单、省时。  相似文献   

10.
为调查分析我国的常规大豆育种材料中是否混入转基因大豆品系,选取706份大豆品系,对可能含有的外源转基因调控元件(CaMV35S启动子、FMV35S启动子和NOS终止子)以及针对我国批准进口的主要耐除草剂转基因大豆转化体(GTS40-3-2和MON89788)成分进行PCR扩增,同时根据MON89788转化体侧翼序列和目...  相似文献   

11.
12.
Summary The effects of the leaves of five plant species, one from each of the generaAmbrosia, Anemone, Eupatorium, Eucalyptus andLantana, on potato tuber moth were investigated under indigenous storage conditions at the Central Potato Research Station, Shillong (1800 m above sea level). Their action was compared with that of a biological insecticide (spores ofBacillus thuringiensis), a chemical insecticide (carbaryl), and an untreated control. The data collected after six months storage on tuber damage, sprout damage and the rotting indicated that the leaves ofLantana aculeata provided most protection to the tubers, reducing damage from over 70% in the check to below 5%, and sprout damage from over 45% to below 3%. Next best wasEucalyptus globulus followed byB. thuringiensis. They may be used on tubers stored for table use or for seed as they had no adverse effect on germination or on the yield of a subsequent crop.  相似文献   

13.
Summary

The efficiency of N fertilizers is usually poor; often less than 50% of the applied N is taken up by the crop. This review focuses on various N fertilizers with respect to the significance of different N loss pathways, namely (i) ammonia volatilization, (ii) dinitrogen and nitrogen oxide emissions, and (iii) nitrate leaching. Further, the significance of biological N immobilization, ammonium fixation and, finally, the impact of nitrate vs. ammonium uptake on crop yield are also discussed. The reviewed literature shows that N fertilizers may differ markedly in their susceptibility to losses. There is, however, considerable scope to improve N efficiency of each N source by proper N management practices.  相似文献   

14.
15.
Summary The in vitro antimicrobial activity of extracts from accessions ofSolanum commersonii Dun. collected in the south of Uruguay was investigated against five microorganisms including the pathogenRalstonia solanacearum. A total of 30 extracts corresponding to organic and aqueous extracts were studied. Interestingly, most of the positive results for growth inhibition were againstR. solanacearum. The extracts were also analyzed for the presence of glycoalkaloids and lectins. Six of the ten aqueous extracts showed lectin presence and a wide variation in the type and amounts of glycoalkaloids, was found. Results indicate that there is no clear relationship between the antimicrobial activity against the five microorganisms screened and the presence or amounts of lectins and glycoalkaloids, traditionally regarded as possible antimicrobial metabolites in theSolanum genus, which indicates the presence of as yet unidentified antimicrobial compounds.  相似文献   

16.
Novel food and non-food uses for sorghum and millets   总被引:4,自引:3,他引:4  
Sorghum and millets have considerable potential in foods and beverages. As they are gluten-free they are suitable for coeliacs. Sorghum is also a potentially important source of nutraceuticals such antioxidant phenolics and cholesterol-lowering waxes. Cakes, cookies, pasta, a parboiled rice-like product and snack foods have been successfully produced from sorghum and, in some cases, millets. Wheat-free sorghum or millet bread remains the main challenge. Additives such as native and pre-gelatinised starches, hydrocolloids, fat, egg and rye pentosans improve bread quality. However, specific volumes are lower than those for wheat bread or gluten-free breads based on pure starches, and in many cases, breads tend to stale faster. Lager and stout beers with sorghum are brewed commercially. Sorghum's high-starch gelatinisation temperature and low beta-amylase activity remain problems with regard to complete substitution of barley malt with sorghum malt . The role of the sorghum endosperm matrix protein and cell wall components in limiting extract is a research focus. Brewing with millets is still at an experimental stage. Sorghum could be important for bioethanol and other bio-industrial products. Bioethanol research has focused on improving the economics of the process through cultivar selection, method development for low-quality grain and pre-processing to recover valuable by-products. Potential by-products such as the kafirin prolamin proteins and the pericarp wax have potential as bioplastic films and coatings for foods, primarily due to their hydrophobicity.  相似文献   

17.
Summary Clones derived from thirty-one different accessions (nineteen of Argentine origin) belonging to eightSolanum species were screened for resistance to infection by potato virus X strain cp (PVX cp) by mechanical inoculation of plantlets that had been micropropagated in vitro. Estimates of PVX multiplication obtained by enzyme linked immunosorbent assay and slot blot nucleic acid hybridization allowed the identification of resistant clones derived from five accessions belonging toS. commersonii S. oplocense, S. sparsipilum andS. tuberosum andigena. Resistant genotypes supported PVX concentrations 5 to 15 times smaller than did the susceptible control cultivar Spunta. Graft inoculation test confirmed the presence of extreme resistance similar to that conferred by the ‘immunity’ gene X1 (also called RXact).  相似文献   

18.
The peer-reviewed marine pharmacology literature from 2009 to 2011 is presented in this review, following the format used in the 1998–2008 reviews of this series. The pharmacology of structurally-characterized compounds isolated from marine animals, algae, fungi and bacteria is discussed in a comprehensive manner. Antibacterial, antifungal, antiprotozoal, antituberculosis, and antiviral pharmacological activities were reported for 102 marine natural products. Additionally, 60 marine compounds were observed to affect the immune and nervous system as well as possess antidiabetic and anti-inflammatory effects. Finally, 68 marine metabolites were shown to interact with a variety of receptors and molecular targets, and thus will probably contribute to multiple pharmacological classes upon further mechanism of action studies. Marine pharmacology during 2009–2011 remained a global enterprise, with researchers from 35 countries, and the United States, contributing to the preclinical pharmacology of 262 marine compounds which are part of the preclinical pharmaceutical pipeline. Continued pharmacological research with marine natural products will contribute to enhance the marine pharmaceutical clinical pipeline, which in 2013 consisted of 17 marine natural products, analogs or derivatives targeting a limited number of disease categories.  相似文献   

19.
Chitosan is considered to be one of the most promising and applicable materials in adsorption applications. The existence of amino and hydroxyl groups in its molecules contributes to many possible adsorption interactions between chitosan and pollutants (dyes, metals, ions, phenols, pharmaceuticals/drugs, pesticides, herbicides, etc.). These functional groups can help in establishing positions for modification. Based on the learning from previously published works in literature, researchers have achieved a modification of chitosan with a number of different functional groups. This work summarizes the published works of the last three years (2012–2014) regarding the modification reactions of chitosans (grafting, cross-linking, etc.) and their application to adsorption of different environmental pollutants (in liquid-phase).  相似文献   

20.
The loss of density and elasticity, the appearance of wrinkles and hyperpigmentation are among the first noticeable signs of skin aging. Beyond UV radiation and oxidative stress, matrix metalloproteinases (MMPs) assume a preponderant role in the process, since their deregulation results in the degradation of most extracellular matrix components. In this survey, four cyanobacteria strains were explored for their capacity to produce secondary metabolites with biotechnological potential for use in anti-aging formulations. Leptolyngbya boryana LEGE 15486 and Cephalothrix lacustris LEGE 15493 from freshwater ecosystems, and Leptolyngbya cf. ectocarpi LEGE 11479 and Nodosilinea nodulosa LEGE 06104 from marine habitats were sequentially extracted with acetone and water, and extracts were analyzed for their toxicity in cell lines with key roles in the skin context (HaCAT, 3T3L1, and hCMEC). The non-toxic extracts were chemically characterized in terms of proteins, carotenoids, phenols, and chlorophyll a, and their anti-aging potential was explored through their ability to scavenge the physiological free radical superoxide anion radical (O2•−), to reduce the activity of the MMPs elastase and hyaluronidase, to inhibit tyrosinase and thus avoid melanin production, and to block UV-B radiation (sun protection factor, SPF). Leptolyngbya species stood out for anti-aging purposes: L. boryana LEGE 15486 presented a remarkable SPF of 19 (at 200 µg/mL), being among the best species regarding O2•− scavenging, (IC50 = 99.50 µg/mL) and also being able to inhibit tyrosinase (IC25 = 784 µg/mL), proving to be promising against UV-induced skin-aging; L. ectocarpi LEGE 11479 was more efficient in inhibiting MMPs (hyaluronidase, IC50 = 863 µg/mL; elastase, IC50 = 391 µg/mL), thus being the choice to retard dermal density loss. Principal component analysis (PCA) of the data allowed the grouping of extracts into three groups, according to their chemical composition; the correlation of carotenoids and chlorophyll a with MMPs activity (p < 0.01), O2•− scavenging with phenolic compounds (p < 0.01), and phycocyanin and allophycocyanin with SPF, pointing to these compounds in particular as responsible for UV-B blockage. This original survey explores, for the first time, the biotechnological potential of these cyanobacteria strains in the field of skin aging, demonstrating the promising, innovative, and multifactorial nature of these microorganisms.  相似文献   

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