Identification of apoptosis regulatory genes using insertional mutagenesis.
This chapter describes a retroviral insertion mutagenesis approach using replication-deficient myeloproliferative sarcoma virus retroviral vectors to identify apoptosis regulatory genes in the interleukin-3-dependent Baf-3 cell line. We describe the retroviral insertion mutagenesis protocol and the selection steps to obtain apoptosis resistant mutants. We also present several methods to isolate the cellular DNA sequences flanking the provirus to identify the gene responsible for the apoptosis-resistant phenotype.
Thomas
Joëlle
J
Laboratory of Molecular and Cellular Biology, Unité de Recherche CNRS/ENS, Lyon, France.
Leverrier
Yann
Y
Mathieu
Anne-Laure
AL
Marvel
Jacqueline
J
D000818:Animals; D017209:Apoptosis / Q000235:genetics* / Q000502:physiology; D002460:Cell Line; D007377:Interleukin-3 / Q000378:metabolism; D051379:Mice; D016254:Mutagenesis, Insertional / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods; D012190:Retroviridae / Q000235:genetics / Q000378:metabolism
D007377:Interleukin-3