| Chapter title |
Detection of In Vivo Mutation in the Pig-a Gene of Mouse Bone Marrow Erythroids
|
|---|---|
| Chapter number | 8 |
| Book title |
Drug Safety Evaluation
|
| Published in |
Methods in molecular biology, January 2017
|
| DOI | 10.1007/978-1-4939-7172-5_8 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-7170-1, 978-1-4939-7172-5
|
| Authors |
Takafumi Kimoto, Daishiro Miura |
| Abstract |
Detection of in vivo mutation is important for evaluating the health risks associated with chemicals. The Pig-a in vivo gene mutation assay has been developed over the last decade for this purpose. Most approaches for the assay, however, measure cells with a Pig-a mutant phenotype in erythrocytes from the peripheral blood, with the mutations causing the phenotype being difficult to determine directly. This chapter describes a procedure for detecting mutations in the Pig-a gene of phenotypically mutant mouse bone marrow erythroids, the precursors of peripheral blood erythrocytes. The strategy for molecular analysis of Pig-a gene mutation includes enrichment of GPI-anchor deficient cells with a cell sorter followed by a cloning and sequencing of Pig-a cDNAs. |
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