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评述了几种常用的体外致突变检测方法及其用于生物样品检测的可行性,有些方法经改进可用于高通量检测生物样品体外致突变性.经典Ames实验受生物样品中组氨酸的影响,易产生假阳性结果,尽管经过修正可以排除组氨酸的干扰,但操作繁琐,不适合高通量检测.基于SOS反应的检测体系避开了组氨酸的影响,且简单易行,适合高通量检测:以β-半乳糖苷酶基因(lacZ)作为报告基因的检测体系灵敏度高,且经过离心洗涤或后培养的方式可降低样品颜色的影响;以绿色荧光蛋白(GFP)基因作为报告基因的检测体系避开了颜色的干扰,但这类方法灵敏度普遍不高,可以寻找信号更强的荧光蛋白以替代GFP;荧光素酶(lux)基因集lacZ和GFP的优点于一身,但检测时需要额外添加辅助因子,限制了其应用.也对单细胞凝胶电泳、tk基因突变实验、染色体损伤检测等方法进行了分析,有些适合生物样品高通量检测,但由于缺少国际通用的标准,很难推广使用.
Several commonly used in vitro mutagenesis detection methods and their feasibility for biological sample detection are reviewed and some of them are improved for high-throughput detection of in vitro mutagenicity of biological samples.A classic Ames experiment is affected by histidine Affect the easy to produce false-positive results, although the modification can rule out the interference of histidine, but the operation is cumbersome, not suitable for high-throughput detection.SOS-based detection system to avoid the influence of histidine, and simple, Suitable for high-throughput detection: the detection system using lacZ gene as a reporter gene has high sensitivity and can reduce the influence of sample color after centrifugation washing or post-culture; and the green fluorescent protein (GFP) gene As a reporter system to avoid the interference of color detection system, but the sensitivity of such methods is generally not high, you can find more signal fluorescent protein to replace GFP; luciferase (lux) gene pool lacZ and GFP advantages in one, However, additional additional cofactors are required for detection, which limits the application of the cofactors.It also analyzes single-cell gel electrophoresis, tk gene mutation test and chromosome damage detection methods, and some suitable organisms Product high-throughput assay, but due to the lack of internationally accepted standards, it is difficult to promote the use.