衣藻中的酸性磷酸酶突变体：通过生化、电泳和遗传分析进行分离与表征
Acid phosphatase mutants in Chlamydomonas: isolation and characterization by biochemical, electrophoretic and genetic analysis.
作者信息
Loppes R, Matagne R F
出版信息
Genetics. 1973 Dec;75(4):593-604. doi: 10.1093/genetics/75.4.593.
Abstract
In order to isolate acid phosphatase mutants in the green alga Chlamydomonas reinhardi, a staining method for detecting the enzyme activity in colonies has been developed. The occurrence of more than one acid phosphatase brought about some difficulty in the selection of mutants. We have, however, found an original method of selection based on the differential heat sensitivity of the enzymes. After treatment of the wild-type strain with N-methyl-N'-nitro-N-nitrosoguanidine, two types of mutants were recovered, then analyzed by biochemical and electrophoretic methods. In the first class of mutants (P(1), P(2), P(3),...) a heat-stable acid phosphatase bound to cellular debris of the crude extract was missing. The mutant P(a), representing the second class of mutations, was lacking a soluble heat-sensitive enzyme. These mutations were genetically different and exhibited mendelian inheritance.
摘要
为了在莱茵衣藻中分离酸性磷酸酶突变体，已开发出一种用于检测菌落中酶活性的染色方法。多种酸性磷酸酶的存在给突变体的筛选带来了一些困难。然而，我们发现了一种基于酶的不同热敏感性的原始筛选方法。用N-甲基-N'-硝基-N-亚硝基胍处理野生型菌株后，获得了两种类型的突变体，然后通过生化和电泳方法进行分析。在第一类突变体（P(1)、P(2)、P(3)……）中，与粗提物细胞碎片结合的热稳定酸性磷酸酶缺失。代表第二类突变的突变体P(a)缺乏一种可溶性热敏感酶。这些突变在遗传上是不同的，表现出孟德尔遗传。
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