苛求芽孢杆菌尿酸酶的表征、应用与其基因克隆

发布时间：2018-01-21 10:07

本文关键词： 苛求芽孢杆菌尿酸酶血清尿酸测定专利尿酸酶法黄嘌呤干扰简并引物PCR 基因克隆　出处：《重庆医科大学》2006年硕士论文　论文类型：学位论文

【摘要】： 测定血清尿酸是临床检验常规工作之一。利用尿酸酶的高专一性进行酶法分析测定血清尿酸含量,即尿酸酶法,是目前常规测定血清尿酸的主要方法。现有尿酸酶法中,过氧化物酶偶联尿酸酶的间接终点平衡法最常用,但此终点平衡法受血清中还原物质和过氧化物,及黄嘌呤等的显著干扰。本实验室在国家自然科学基金(30200266)资助下,建立了血清尿酸含量测定的专利方法(zl 03135649.4),此专利方法对血清中常见干扰物质不敏感;与其它尿酸酶法相比,相同效率下其成本最低,线性范围最宽,对黄嘌呤抗性最强。已报道尿酸酶法中,测定尿酸酶作用下紫外吸收总变化的直接终点平衡法对常见干扰抗性最强。本实验室的专利方法与直接终点平衡法一致,但与最常用的过氧化物酶偶联尿酸酶间接终点平衡法不一致。可见此专利方法可行且可靠。但此专利方法需要K_m较高且对黄嘌呤抑制不敏感的工具尿酸酶。目前市售尿酸酶K_m都较低且对黄嘌呤敏感。另一方面,尿酸酶也是当前治疗高尿酸血症相关疾病的理想药物。苛求芽孢杆菌依赖于高浓度尿酸而生长,其可表达高比活性的胞内和胞外尿酸酶;此类尿酸酶K_m较高,最接近我们专利方法要求;但至今未见关于此物种的任何基因序列信息报道。为获得廉价且符合要求尿酸酶以促进测定血清
[Abstract]:Determination of serum uric acid is one of the routine tasks of clinical examination. The indirect end point equilibrium method of peroxidase conjugated uricase is the most commonly used, but this end point equilibrium method is by reducing substance and peroxide in serum. And the significant interference of xanthine. The patent method for the determination of serum uric acid was established in our laboratory with the aid of the National Natural Science Foundation of China (30200266). The patent method for determination of serum uric acid is zl03135649.4). The patented method is insensitive to common interfering substances in serum. Compared with other uric acidase methods, it has the lowest cost, the widest linear range and the strongest resistance to xanthine at the same efficiency. The direct end-point equilibrium method for the determination of total ultraviolet absorption under the action of uric acid enzyme is the most resistant to common interference. The patent method in our laboratory is consistent with the direct end-point equilibrium method. However, it is not consistent with the most commonly used indirect end-point equilibrium method for peroxidase conjugated uric acidase. It can be seen that the patented method is feasible and reliable. However, the patented method requires uric acid enzyme with higher km and less sensitivity to xanthine inhibition. At present, uric acidase K _ m is low and sensitive to xanthine, on the other hand. Urease is also an ideal drug for the treatment of hyperuricemia related diseases. Bacillus causticus is dependent on high concentration of uric acid and can express high specific activity of intracellular and extracellular uric acidase. This kind of uric acid enzyme K _ S _ m is relatively high and closest to our patented method requirements; However, no information on the gene sequence of the species has been reported. To obtain cheap and satisfactory uric acid enzymes to facilitate the determination of serum
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R378

【相似文献】