RESUMO
In the production of novel biological products, plasmids are often engineered into delivery vectors for target genes, which can be used directly as vaccines or as intermediate products for gene/cell therapy. Plasmid DNA exists in several topological forms such as supercoiled, linear, and open circular. As supercoiled plasmid shows the highest efficiency in transfecting eukaryotic cells, the content of supercoiled plasmids becomes an important indicator of plasmid quality. CGE is an effective analysis method for separating different topological structures of plasmids. For the purpose of providing plasmid manufacturers and regulatory agencies with an efficient and readily used tool for monitoring the quality of plasmids, this article identifies the optimal separation and detection conditions of CGE, presents a platform-based plasmid analytical method, and uses plasmid of different sizes to verify the feasibility of this method. In terms of detector, the LIF detector has obvious advantages over the ultraviolet detector in sensitivity and resolution. Using the optimal CE condition (10× gel buffer), baseline separation of different topological forms and impurities can be achieved for different plasmid sizes (5.9, 7.8, 15.4 kb). In addition, 6.5 kb plasmid was used to compare the different separation technologies such as CGE-LIF, ion exchange chromatography and agarose gel electrophoresis. The result shows that CGE-LIF can provide better resolution and quantitation accuracy than ion exchange chromatography and agarose gel electrophoresis. CGE-LIF, as a quick and convenient method to separate and quantify plasmids, has the advantages of high sensitivity, high resolution, and high quantitative accuracy. Therefore, it is ideal for analysis of plasmids with different sizes, and it can also be used as a platform method for manufacturers and regulatory agencies to monitor the purity and stability of plasmids.
Assuntos
Eletroforese Capilar , Cromatografia por Troca Iônica/métodos , Eletroforese em Gel de Ágar/métodos , Eletroforese Capilar/métodos , Plasmídeos/genética , Isoformas de ProteínasRESUMO
Temperature-dependent dual fluorescence and switchable circularly polarized luminescence (CPL) are two highly pursued but challenging properties for small organic molecules (SOMs). We herein disclose a triarylborane π-system based on a 2,2'-diamino-6,6'-diboryl-1,1'-binaphthyl scaffold that can serve as a versatile building block for achieving these two properties by simply choosing different amino groups. BNMe2 -BNaph with less bulky dimethylamino groups displays temperature-dependent dual fluorescence, and can thus be used as a highly sensitive ratiometric fluorescence thermometer. On the other hand, BNPh2 -BNaph with bulky diphenylamino groups exhibits intense fluorescence in both solution and in the solid state. A change of solvent from nonpolar cyclohexane to highly polar MeCN not only shifts the CPL position to much longer wavelength but also inverts the CPL sign. In addition, the complexation of BNPh2 -BNaph with fluoride greatly enhances the CPL intensity.
RESUMO
OBJECTIVE: To identify the pathogen of Bletilla striata pseudobulb rot and provide foundation for formulation effective control measures. METHODS: Surveyed the pathogenesis regularity and disease symptom of Bletilla striata pseudobulb rot,and conducted the pathogen separation, identification,vaccination and research on its biological characteristics. RESULTS: The conditions for mycelium growth were as follows: the temperature ranged from 5 to 35 degrees C, and the optimum temperature ranged from 25 to 30 degrees C. The pH ranged from 3 to 10,and the optimum pH ranged from 6 to 7. The germination rate of conidia was high when the relative humidity was 100%. The lethal temperature was 55 degrees C for 10 min. Illumination had little influence on the germ growth and development. CONCLUSION: The results showed that the pathogen was Fusarium oxysporum.
