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Hu, Bo,Liang, Minjian,Hong, Guoqiang,Li, Zhaoxia,Zhu, Zhenyu,Li, Lin Korean Society for Biochemistry and Molecular Biol 2005 Journal of biochemistry and molecular biology Vol.38 No.6
Antibody to hepatitis B surface antigen (HBsAb) is the important serological marker of the hepatitis B virus (HBV) infection. Conventionally, the hepatitis B surface antigen (HBsAg) obtained from the plasma of HBV carriers is used as the diagnostic antigen for detection of HBsAb. This blood-origin antigen has some disadvantages involved in high cost, over-elaborate preparation, risk of infection, et al. In an attempt to explore the suitable recombinant HBsAg for the diagnostic purpose, the HBV S gene was expressed in Pichia pastoris and the product was applied for detection of HBsAb. Hepatitis B virus S gene was inserted into the yeast vector and the expressed product was analyzed by sodium dodecyl sulphate polyacrolamide gel electrophoresis (SDS-PAGE), immunoblot, electronic microscope and enzyme linked immunosorbent assay (ELISA). The preparations of synthesized S protein were applied to detect HBsAb by sandwich ELISA. The S gene encoding the 226 amino acid of HBsAg carrying ahexa-histidine tag at C terminus was successfully expressed in Pichia pastoris. The His-Tagged S protein in this strain was expressed at a level of about 14.5% of total cell protein. Immunoblot showed the recombinant HBsAg recognized by monoclonal HBsAb and there was no cross reaction between all proteins from the host and normal sera. HBsAb detection indicated that the sensitivity reached 10 mIu (micro international unit)/ml and the specificity was 100% with HBsAb standard of National Center for Clinical Laboratories. A total of 293 random sera were assayed using recombinant S protein and a commercial HBsAb ELISA kit (produced by blood-origin HBsAg), 35 HBsAb positive sera and 258 HBsAb negative sera were examined. The same results were obtained with two different reagents and there was no significant difference in the value of S/CO between the two reagents. The recombinant HBV S protein with good immunoreactivity and specificity was successfully expressed in Pichia pastoris. The reagent for HBsAb detection prepared by Pichia pastoris-derived S protein showed high sensitivity and specificity for detection of HBsAb standard. And a good correlation was obtained between the reagent produced by recombinant S protein and commercial kit produced by blood-origin HBsAg in random samples.
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Chen, Yujie,Zhao, Zhongzhen,Chen, Hubiao,Brand, Eric,Yi, Tao,Qin, Minjian,Liang, Zhitao The Korean Society of Ginseng 2017 Journal of Ginseng Research Vol.41 No.1
Background: Asian ginseng and American ginseng are functional foods that share a close genetic relationship and are well-known worldwide. This article aims to investigate the correlation between morphological characteristics and the inherent quality of Asian and American ginsengs. Methods: In this study, an ultra-HPLC-quadrupole/time-of-flight MS (UHPLC-Q/TOF-MS) method was established for the quantitative analysis of 45 ginseng samples. The method developed for determination was precise and accurate. Results: The results showed that Asian ginseng samples with the same growing time (with the same or similar number of stem scars) that had a thinner main root, a longer rhizome and more branch roots contained greater amounts of ginsenosides. For American ginseng, two tendencies were observed in the relationship between the diameter of the main root and contents of ginsenosides. One tendency was that samples with thinner main roots tended to contain higher levels of ginsenosides, which was observed in the samples sold under the commercial name pao-shen. Another tendency was that samples with thicker main roots contained higher contents of ginsenosides, which was observed in the samples sold under the commercial name pao-mian, as well as in samples of American ginseng cultivated in Jilin, China. Conclusion: An approach using ultra-HPLC-quadrupole/time-of-flight MS was successfully established to link morphology and active components for evaluating the quality of Asian and American ginsengs. Clear correlation between visible morphological features and quality of Asian and American ginsengs was found. People can see the difference; this means consumers and vendors can evaluate ginseng by themselves.
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