TY - JOUR
T1 - Cloning and expression of cDNA for rat heme oxygenase
AU - Shibahara, S.
AU - Muller, R.
AU - Taguchi, H.
AU - Yoshida, T.
PY - 1985
Y1 - 1985
N2 - Two cDNA clones for rat heme oxygenase have been isolated from a rat spleen cDNA library in λgt11 by immunological screening using a specific polyclonal antibody. One of these clones had an insert of 1530 nucleotides that contains the entire protein-coding region. To confirm that the isolated cDNA encodes heme oxygenase, we transfected monkey kidney cells (COS-7) with the cDNA carried in a simian virus 40 vector. The heme oxygenase was highly expressed in endoplasmic reticulum of transfected cells. The nucleotide sequence of the cloned cDNA was determined and the priamry structure of heme oxygenase was deduced. Heme oxygenase is composed of 289 amino acids and has one hydrophobic segment at its carboxyl terminus, which is probably important for the insertion of heme oxygenase into endoplasmic reticulum. The cloned cDNA was used to analyze the induction of heme oxygenase in rat liver by treatment with Cocl2 or with hemin. RNA blot analysis showed that both CoCl2 and hemin increased the amount of hybridizable mRNA, suggesting that these substances may act at the transcriptional level to increase the amount of heme oxygenase.
AB - Two cDNA clones for rat heme oxygenase have been isolated from a rat spleen cDNA library in λgt11 by immunological screening using a specific polyclonal antibody. One of these clones had an insert of 1530 nucleotides that contains the entire protein-coding region. To confirm that the isolated cDNA encodes heme oxygenase, we transfected monkey kidney cells (COS-7) with the cDNA carried in a simian virus 40 vector. The heme oxygenase was highly expressed in endoplasmic reticulum of transfected cells. The nucleotide sequence of the cloned cDNA was determined and the priamry structure of heme oxygenase was deduced. Heme oxygenase is composed of 289 amino acids and has one hydrophobic segment at its carboxyl terminus, which is probably important for the insertion of heme oxygenase into endoplasmic reticulum. The cloned cDNA was used to analyze the induction of heme oxygenase in rat liver by treatment with Cocl2 or with hemin. RNA blot analysis showed that both CoCl2 and hemin increased the amount of hybridizable mRNA, suggesting that these substances may act at the transcriptional level to increase the amount of heme oxygenase.
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U2 - 10.1073/pnas.82.23.7865
DO - 10.1073/pnas.82.23.7865
M3 - Article
C2 - 3865203
AN - SCOPUS:0022367081
VL - 82
SP - 7865
EP - 7869
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
SN - 0027-8424
IS - 23
ER -