The In Vivo Pig-a Gene Mutation Assay
The In Vivo Pig-a Gene Mutation Assay
复制标题
猪体内基因突变检测
DOI：
10.3123/jemsge.2014.027
复制
发表时间：
2014
影响因子：
1.7
通讯作者：
D. Miura
中科院分区：
文献类型：
--
作者：
D. Miura
磷脂酰肌醇聚糖锚定生物合成，A类基因（啮齿动物中的猪-A，人类中的猪-A）编码n -乙酰氨基葡萄糖转移酶复合物的催化亚基，该复合物参与糖基磷脂酰肌醇（GPI）锚定合成的早期步骤，GPI锚定将特定的蛋白质标记物系在各种类型细胞的表面。两种不同的策略猪-a基因突变检测使用外周血已经开发，一个使用流式细胞术和另一个使用有限稀释克隆。利用细菌原毒素（原气溶素）作为选择性试剂进行有限稀释克隆试验是资源密集型的。对于突变频率的常规分析，采用针对gpi锚定标记物的荧光标记抗体的流式细胞术程序（例如，大鼠红细胞的抗cd59，小鼠红细胞的抗cd24）是首选的突变频率常规分析方法。然而，克隆试验的优势在于对突变基因型的深入分析（突变谱分析）。流式细胞术猪-a基因突变试验的特点，即诱导猪-a基因突变频率具有持久性和分剂量化学物质的作用具有可加性，使网织红细胞和总红细胞的试验成为开发体内详细致突变性数据的有吸引力的可能性。尽管必须收集大量有关分析性能的信息，但在过去几年中，实现这一目标的进展很快，并且已经启动并正在进行多实验室试验。
The phosphatidylinositol glycan anchor biosynthesis, Class A gene (Pig-a in rodents, PIG-A in humans) codes for a catalytic subunit of the N-acetylglucosamine transferase complex that is involved in an early step of glycosylphosphatidyl inositol (GPI) anchor synthesis, and GPI anchors tether specific protein markers to the surface of various types of cells. Two distinct strategies for Pig-a gene mutation assay used peripheral blood have been developed, one using flow cytometry and the other using limiting-dilution cloning. The limiting-dilution cloning assay using bacterial protoxin, proaerolysin, as a selective agent is resource intensive. For routine analysis of mutant frequency, the flow cytometric procedures employing fluorescently labeled antibodies against GPI-anchored markers (e.g., anti-CD59 for rat red blood cells, anti-CD24 for mouse red blood cells) are preferred for routine analysis of mutant frequency. The advantage of the cloning assay, however, is in-depth analyses of the mutant genotype (mutational spectra analysis). The characteristics of the flow cytometric Pig-a gene mutation assays, i.e., induced Pig-a mutant frequencies were persistence and the effect of split doses of chemicals were additive, make the assay on reticulocytes and total red blood cells an attractive possibility for developing detailed mutagenicity data in vivo. Although a large amount of information must be gathered on assay performance, progress toward the goal in the last few years has been rapid, and multi-laboratory trial has been initiated and ongoing.
影响因子：
3.8
作者：
Phonethepswath, Souk;Franklin, Dean;Dertinger, Stephen D.
通讯作者：
Dertinger, Stephen D.
DOI：
10.1073/pnas.96.9.5209
发表时间：
1999-04-27
影响因子：
11.1
作者：
Araten, DJ;Nafa, K;Luzzatto, L
通讯作者：
Luzzatto, L