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Title: Stable expression of eukaryotic p450 cDNA in mammalian cell lines
Author: Doostdar, Hamed
Awarding Body: University of Aberdeen
Current Institution: University of Aberdeen
Date of Award: 1992
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The P450 profile of HepG2 cells was studied using specific enzyme assays and well characterised monoclonal and polyclonal antibodies. These cells were shown to endogenously express cytochrome P450 1A1, P450 1A2 and P450 3A5 when the cells were treated with benz(a)anthracene or rifampicin respectively. HepG2 cells were adapted to grow in Williams' E medium supplemented with only 5% (v/v) fetal calf serum. These cells showed a significantly higher concentration of P450 1A2 protein when treated with benz(a)anthracene and could be detected by P450 1A2 specific antibodies on Western blots. Stable cell lines of HepG2 and Cos-7 cells capable of continuous expression of the Epstein-Barr virus nuclear antigen-1 (EBNA-1) were constructed. P450 2A6 cDNA was successfully expressed in the EBNA-1 transformed Cos-7 cell line for over 6 weeks. The rate of expression of this cDNA and the enzyme activity of the resulting protein was very low. No stable HepG2 EBNA-1 transformed cells capable of P450 2A6 expression was obtained.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available