Establishment of immunochemical determination of human cholesterol 7 alpha-hydroxylase

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A fusion protein was prepared from an in-frame fusion gene containing the cDNA for human cholesterol 7alpha-hydroxylase near the 3' terminus of the lacZ gene of Escherichia coli and monoclonal antibody angainst human cholesterol 7alpha-hydroxylase was established. Using this antibody, immunochemical determination was established.To clarify the enzyme level of cholesterol 7alpha-hydroxylase during liver regeneration, we determined the enzyme level after hepatectomy in the rats. We also determined mRNA level and enzyme activity. 12hrs after hepatectomy, the level of mRNA,enzyme activity and enzyme protein mass decreased in parallel and gradually returned to normal level. These results suggest that this enzyme protein decreased during cell division and is under pretranslational regulation. Now we are preparing the manuscript concerning the species difference of gene expression of cholesterol 7alpha-hydroxylase after bile duct ligation in the rats and rabbits.In the course of pursuing the project, we have investigated the enzymes, 3beta-hydroxy-DELTA^5-C_<27>-steroid dehydrogenase and 25-hydroxyvitamin D_3 1alpha-hydroxylase, which are related to the cholesterol 7alpha-hydroxylase and published in the journals.