Expression efficiency of the human thrombomodulin-encoding gene in various vector and host systems

Gene
J H LinJ Morser

Abstract

Expression systems were developed for evaluating recombinant human thrombomodulin (TM) production in different host cell lines by investigating the performance of five mammalian expression vectors. The expression vectors were constructed so that they contain multiple monocistronic gene cassettes which include a gene encoding a dominant selectable marker, HyR (hygromycin B phosphotransferase), under the regulation of the thymidine kinase promoter, the target gene which encodes a truncated human re-TM under the regulation of various promoters, an amplifiable gene (Dhfr) encoding murine dihydrofolate reductase under the regulation of either the SV40 early or late promoter along with the SV40 enhancer and the SV40 ori. We tested the performance of the five expression vectors in human embryonic kidney cells (HEK293), baby hamster kidney cells (BHK), human melanoma cells (CHL-1) and Dhfr- Chinese hamster ovary cells (CHO/Dhfr-). We found that the efficiency of DNA uptake, transient expression and stable expression of the different expression vectors were all cell-line dependent. However, the myeloproliferative sarcoma virus (MPSV) LTR promoter consistently showed higher expression levels in all cell lines, particularly in HEK293 cell...Continue Reading

References

Jul 1, 1980·Proceedings of the National Academy of Sciences of the United States of America·G Urlaub, L A Chasin

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Citations

May 22, 2002·Biochimica Et Biophysica Acta·Ralph MelcherAndrej Hasilik
Oct 20, 2005·Journal of Bioscience and Bioengineering·Masahiro KawaharaTeruyuki Nagamune
Sep 1, 1995·Science·J FengJ Yu
Dec 17, 2009·BMC Biotechnology·Kristina NehlsenDagmar Wirth
Apr 1, 2000·The Journal of Biological Chemistry·M M OuelletteW E Wright

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