Use of bleomycin- and heat shock-induced calreticulin promoter for construction of a mammalian expression vector.
Addition of bleomycin (Bm) to an NIH/3T3 cell culture induced the overproduction of four cellular proteins [Kumagai and Sugiyama (1998) J. Biochem. 124, 835-841]. The two proteins were identified on N-terminal amino acid sequence analysis as calreticulin and mitochondrial matrix protein P1, which are known as heat shock proteins, respectively. In this study, we cloned the calreticulin promoter region from the genomic DNA of NIH/3T3 cells and observed that heat shock treatment at 42 degrees C or the addition of Bm to the cell culture caused overexpression of the luciferase gene controlled by the cloned calreticulin promoter. This suggests that Bm induces the transcriptional activation of stress-heat shock genes. We constructed an expression vector for mammalian cells, which is controlled by the calreticulin promoter.
Elmileik
H
H
Department of Pediatrics, School of Medicine, and Institute of Pharmaceutical Sciences, Faculty of Medicine, Hiroshima University, Kasumi Minami-ku, Hiroshima 734-8551, Japan. sugi@hiroshima-u.ac.jp
Kumagai
T
T
Berengena
M
M
Ueda
K
K
Sugiyama
M
M
D016475:3T3 Cells; D000818:Animals; D001761:Bleomycin / Q000378:metabolism / Q000494:pharmacology*; D002135:Calcium-Binding Proteins / Q000096:biosynthesis / Q000187:drug effects / Q000235:genetics*; D037282:Calreticulin; D018834:Chaperonin 60 / Q000096:biosynthesis / Q000187:drug effects / Q000235:genetics*; D003001:Cloning, Molecular; D005822:Genetic Vectors / Q000235:genetics*; D018869:Heat-Shock Response / Q000528:radiation effects; D008156:Luciferases / Q000187:drug effects / Q000235:genetics / Q000528:radiation effects; D051379:Mice; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012261:Ribonucleoproteins / Q000096:biosynthesis / Q000187:drug effects / Q000235:genetics*; D015533:Transcriptional Activation / Q000187:drug effects / Q000235:genetics
D002135:Calcium-Binding Proteins; D037282:Calreticulin; D018834:Chaperonin 60; D012261:Ribonucleoproteins; D001761:Bleomycin; D008156:Luciferases