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High-level Expression and Characterization of the Human Interleukin-10 in the Milk of Transgenic Mice

  • Zneng, Z. Y.;B. H. Sohn;K. B. Oh;W. J. Shin;Y. M. Han;Lee, K. K.
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.46-46
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    • 2003
  • Interleukin-10 (IL-10) is a homodimeric protein with a wide spectrum of anti-inflammatory and immune activities. It inhibits cytokine production and expression of immune surface molecules in various cell types. The transgenic mice carrying the human IL-10 gene in conjunction with the bovine $\beta$-casein promoter produced the human IL-10 in milk during lactation. Transgenic mice were generated using a standard method as described previously. To screen transgenic mice, PCR was carried out using chromosomal DNA extracted from tail or toe tissues with a primer set. In this study, stability of germ line transmission and expression of IL-10 gene integrated into host chromosome were monitored up to generation F15 of a transgenic line. When female mouse of generation F9 was crossbred with normal male, generation F9 to F15 mice showed similar transmission rates (66.0$\pm$20.13%, 61.5$\pm$16.66%, 41.1$\pm$8.40%, 40.7$\pm$20.34%, 61.3$\pm$10.75%, 49.2$\pm$18.82%, and 43.8$\pm$25.91%, respectively), implying that the IL-10 gene can be transmitted stably up to long term generation in the transgenic mice. For ELISA analysis, IL-10 expression levels were determined with an hIL-10 ELISA and a mIL-10 ELISA kit in accordance with the supplier's protocol. Expression levels of human IL-10 from milk of generation F9 to F13 mice were 3.6$\pm$1.20 mg/ml, 4.2$\pm$0.93 mg/ml, 5.7$\pm$1.46 mg/ml, 6.3$\pm$3.46 mg/ml, and 6.8$\pm$4.52 mg/ml, respectively. These expression levels are higher than in generation F1 (1.6 mg/ml) mice. We concluded that transgenic mice faithfully passed the transgene on their progeny and successively secreted target proteins into their milk through several generations, although there was a little fluctuation in the transmission frequency and expression level between the generations.

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Effects or $H_2O$ Fraction or Dioscorea japonica Thunb with selenium on plasma Glucose and Lipid Metabolism in Streptozotocin Induced Diabetic Rats (당뇨 흰쥐에서의 참마 $H_2O$ 분획물과 Selenium 보충시 혈당 및 지질대사에 미치는 영향)

  • 김명화
    • Journal of Nutrition and Health
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    • v.31 no.9
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    • pp.1377-1384
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    • 1998
  • The purpose of this study was to investigate the effect of H$_2$O fraction of Dioscorea japonica Thunb(DJT) with selenium(Se) treatment on blood glucose and lipid metabolism in streptozotocin(STZ)-induced diabetic rats. Male Sprague-Dawley rats(180-220g) weighing were divided into five groups, that is one normal group and four diabetic experimental groups : the STZ-control group, the DJT group, the DJT-Se group and the Se group. Diabetes mellitus was induced in the male rats by injection of STZ into the tail vein at a dose of 45mg/kg B.W. The H,0 fraction of DJT(500mg/kg) given orally were administered for 14 days. The Se treated group were fed a AIN-76 recommendation diet mixed with Na$_2$SeO$_3$(2mg/kg diet), which was prepared fresh daily. The body weight and food intake was monitored daily and plasma levels of glucose, insulin, hematocrit and protein were determined. The plasma concentrations of cholesterol, HDL-cholesterol, triglyceride and fire fatty acid were measured. The activities of aminotrans ferase were analysed. The body weight gain was shown to be significantly higher in the normal group than all diabetic groups. The blood glucose levels of the DIT-Se group was significantly lower compared to those of the experimental groups. The administration of H$_2$O fraction of DJT and selenium showed an increase in plasma protein concentrations. The plasma cholesterol levels of all STZ-groups were not significantly different and HDL-cholesterol levels were increased in the diabetic experimental groups fed on H$_2$O fraction of DJT or Se supplementation. Plasma triglyceride levels were lower in the DJT-Se and Se group than in the STZ-control group. free fatty acid levels were not significantly differ among STZ-control groups. STZ treatment increased aminotransferase activity and that of DJT group was highest. In conclusion, the data from the present experiments indicate that the treatment of the H$_2$O fraction of DJT with selenium showed a synergistic effect and the two can have an influence on hyperg1ycemia and lipid metabolites when administered together. (Korean J Nutrition 31(9) : 1377-1384, 1998)

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