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1.
Chinese Journal of Preventive Medicine ; (12): 721-725, 2010.
Article in Chinese | WPRIM | ID: wpr-291508

ABSTRACT

<p><b>OBJECTIVE</b>To achieve secretory and extracellular production of recombinant dengue virus serotypes I-IV envelope glycoprotein domain III (DENV-1-4 EDIII) in Pichia pastoris.</p><p><b>METHODS</b>EDIII genes of DENVI-IV were amplified and cloned into vector pPIC9K, respectively. These recombinant plasmids were then linearized and transferred into Pichia pastoris strain GS115. Clones highly produced in 4.0 mg/ml G418 were amplified and induced by methanol to achieve the secreted recombinant proteins. Ni-NTA agarose beads were used for purification, while SDS-PAGE and Western blotting were used for identification.</p><p><b>RESULTS</b>The recombinant plasmids pPIC9K-DENV-1-4 EDIII were constructed and successfully transferred into Pichia pastoris strain GS115. The recombinant EDIII proteins were expressed in a secretory way with the molecular weight about 12 × 10(3) and specifically identified by anti-His monoclonal antibody and anti-DENVI-IV mice sera.</p><p><b>CONCLUSION</b>DENVI-IV EDIII proteins are successfully achieved from Pichia pastoris expression system and could be used for development of dengue vaccines, diagnostic reagents and study of biological function of the E protein.</p>


Subject(s)
Dengue Virus , Genetics , Genetic Vectors , Pichia , Metabolism , Recombinant Proteins , Genetics , Viral Envelope Proteins , Bodily Secretions
2.
China Journal of Chinese Materia Medica ; (24): 175-177, 2005.
Article in Chinese | WPRIM | ID: wpr-276623

ABSTRACT

<p><b>OBJECTIVE</b>To study the characteristic features of Desmodium gyrans in order to provide a basis for rational exploitation and utilization of the herb.</p><p><b>METHOD</b>Samples of the title plant were collected, the microscopic features of cross sections and powders were studied. TLC profiles and UV absorption of the plant extract were examined.</p><p><b>RESULT</b>Calcium oxalate crystals were found in cells of transverse sections. Nonglandular hairs were observed on leaf surfaces. Characteristic peaks in the UV spectrum were identified.</p><p><b>CONCLUSION</b>The distinct characteristic features revealed in this studies can serve as evidence for the identification of D. gyrans.</p>


Subject(s)
Chromatography, Thin Layer , Fabaceae , Chemistry , Cell Biology , Pharmacognosy , Plant Leaves , Chemistry , Cell Biology , Plant Stems , Chemistry , Cell Biology , Plants, Medicinal , Chemistry , Cell Biology , Powders , Quality Control , Spectrophotometry, Ultraviolet
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