摘要
采用“拍板技术”(Flap technique),自制超薄层(0.2mm)聚丙烯酰胺凝胶板.检测了九株Ceftazidime耐药细菌的β-内酰胺酶,并与标准染色体介导的β-内酰胺酶进行比较。除R27853具有两条酶带外,其余均为一条酶条。R1006,R8701,R884三种酶的等电点(pI)与标准酶D-31一致,pI为8.6;R222,R1029两种酶的pI与标准酶P99吻合,pI为8.0。同时,还发现了pI为8.2的R27853和pI为8.4的R471、R855、R875四种酶,目前尚未见文献报道。这一超薄层凝胶技术具有操作简便、快速、灵敏、省时,便于同步比较多个样品及实验结果更易保存等优点,可用作β-内酰胺酶的鉴定与分类,以及进行蛋白质成份分析的方法。
Ultrathin polyacrylamide gels(0.2mm) were prepared by the new 'flap technique'.Through ultrathin-layer analytical isoeletcric focusing electrophorsi s, the β-lactamase focusing patterns of each strain (9 Ceftazidime-resistant strains, in which 7 E.cloacae, 2 P. aeruginosa) consisted of a single band except that enzyme R27853 had 2 bands. The isoelectric points (pI) of β-lactamases R10C6, R8701 and R884 were the same as that of standard chromosamal-mediated β-lactamase D-31 (pI = 8.6), the pis of β-lactamases R222 and R1029 were similar to that of standard chromosamal-mediated β-lactamase P-99 (pI = 8.0). In addition,β-lactamase R27853 (pI = 8.2) and β-lactamases R471, R585 and R875(pI = 8.4)respetcively may be new chromosamal β-lacta mases.
The experiments suggested that the ultrathin-layer isoelectric focusing electrophore-sis characterized by its high resolution, high speed, versatility and simplicity of operation, and recommends itself as a good analytical technique for research study and routine applications.
出处
《中国抗生素杂志》
CAS
CSCD
北大核心
1990年第3期184-187,共4页
Chinese Journal of Antibiotics
关键词
内酰胺酶
超薄层
等电聚焦
Ultrathin-layer isoelectric focusinge ecltrophoresis
β-lactamase.
