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CYP2C9*58型新突变体的体外酶学活性研究 被引量:1
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作者 戴大鹏 李传保 +3 位作者 王双虎 耿培武 胡国新 蔡剑平 《医学研究杂志》 2013年第11期58-61,共4页
目的对CYP2C9基因新突变体(CYP2C9*58型)开展体外酶学功能研究,明确其代谢活性与野生型的相关性。方法以CYP2C9基因cDNA为模板,通过定点诱变方法获得CYP2C9各变异体的cDNA全长,用以构建昆虫表达载体。利用杆状病毒包装试剂盒包装昆虫病... 目的对CYP2C9基因新突变体(CYP2C9*58型)开展体外酶学功能研究,明确其代谢活性与野生型的相关性。方法以CYP2C9基因cDNA为模板,通过定点诱变方法获得CYP2C9各变异体的cDNA全长,用以构建昆虫表达载体。利用杆状病毒包装试剂盒包装昆虫病毒,侵染sf21昆虫细胞后获得高效表达CYP2C9各型蛋白的微粒体。以甲苯磺丁脲为探针底物药,利用获得的微粒体体外测定各变异体的最大反应速率V max和米氏常数K m,评价其主要酶促动力学特性。结果成功构建了CYP2C9*1、CYP2C9*2、CYP2C9*3和CYP2C9*58型4种CYP2C9昆虫表达载体,Western blot证实该载体可用于稳定表达相应的CYP2C9突变体。CYP2C9*2、CYP2C9*3和CYP2C9*58型变异体的体外酶学活性分别为野生型CYP2C9*1的91.6%,13.5%和23.1%。结论 CYP2C9*58型的酶学活性较野生型明显降低,接近于典型缺陷型突变体CYP2C9*3型,提示携带此突变型的患者在服用经由CYP2C9代谢的相关药物时,药物代谢速度较野生型携带者会有一定程度的降低。 展开更多
关键词 CYP2C9 新突变体体外酶活性研究 慢代谢
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评价一种新CYP2C9突变型对药物代谢的影响
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作者 亢奋恒 张博文 +2 位作者 戴大鹏 李传宝 蔡剑平 《医学研究杂志》 2019年第6期54-59,共6页
目的在体外表达细胞色素P2C9(32T> C),构建酶促反应体系,检测其对甲苯磺丁脲(tolbutamide)的代谢活性.方法利用定向诱变技术,以细胞色素P2C9野生型的互补DNA为反义链,获得细胞色素P2C9*3以及细胞色素P2C9(32T>C)的complementary D... 目的在体外表达细胞色素P2C9(32T> C),构建酶促反应体系,检测其对甲苯磺丁脲(tolbutamide)的代谢活性.方法利用定向诱变技术,以细胞色素P2C9野生型的互补DNA为反义链,获得细胞色素P2C9*3以及细胞色素P2C9(32T>C)的complementary DNA,利用杆状病毒穿梭载体bacmid抽提试剂盒获得bacmid,转染伪黏虫卵巢细胞,充分表达后提取微粒体蛋白质,并用免疫印迹法检测其表达量.在体外条件下,测定各型细胞色素P2C9催化甲苯磺丁脲代谢的最大反应速率Vm和米氏常数Km,评价新突变体的催化功能.结果利用昆虫表达系统表达获得细胞色素P2C9 * 1、细胞色素P2C9*3和细胞色素P2C9(32T> C)3种微粒体蛋白质.免疫印迹结果显示,新变异型蛋白表达水平接近于野生型.体外代谢实验结果显示,细胞色素P2C9(32T> C)针对甲苯磺丁脲的体外清除率(Vm/Km)明显低于野生型,体外酶学活性为野生型的12.40%.结论在体外条件下,细胞色素P2C9(32T> C)对甲苯磺丁脲的代谢活性明显下降,提示携带有该基因型的患者服用细胞色素P2 C9的底物药时,体内代谢速率可能会有不同程度的降低,用药前需考虑调整剂量. 展开更多
关键词 细胞色素P2C9 新突变体 体外研究
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Economic Impact of Mutant Cassava (Manihot Species) for Agricultural Improvement in Ghana
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作者 Kwamina Ewur Banson Harry Amoatey Joe Frederick Cobbinah 《Journal of Agricultural Science and Technology(B)》 2014年第4期272-280,共9页
The importance of agricultural innovations for raising the living standards of farmers has received substantial attention from the economic and political communities. In Ghana, cassava serves as an important source of... The importance of agricultural innovations for raising the living standards of farmers has received substantial attention from the economic and political communities. In Ghana, cassava serves as an important source of calorie and cash crop. Production constraints such as the cassava mosaic disease caused by virus have limit farmers access to productivity potentials and marketing. Breeding efforts attempting to address this constraints have had some success such as the mutant Tech Bankye virus resistant variety. This research, therefore, analysed the impact of some production input on mutant cassava productivity growth in Ghana with the use of Cobb-Douglas production function. Results show that farmers cultivate the mutant variety in addition to other traditional varieties to averse risking in marketing and income. The average cost of production of the mutant variety is higher compared to that of the traditional variety. Eighty percent of the farmers complained difficulty in accessing market for the mutant variety because it could not replace the traditional variety in terms of its texture for the local native "fufu" production and "ampesi" and also deteriorate quickly in storage. However, for industrial production of "gari" and starch, the mutant is the best. Both the mutant cassava and the traditional varieties showed increasing returns to scale, however, the returns to scale of the mutant variety were lower compared to that of the traditional variety. Also, the opportunity cost of forgoing the traditional varieties to cultivating the mutant variety was very high making adopted farmers hesitant to continue it cultivation. 展开更多
关键词 Technology adoption market production cost production function mutant variety opportunity cost returns to scale agricultural productivity.
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