Efficient expression of pro-urokinase by human lymphoblastoid Namalwa KJM-1 cells using Moloney retroviral promoter |
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Authors: | Mitsuo Satoh Hiromasa Miyaji Tatsunari Nishi Tamio Mizukami Seiji Sato Seiga Itoh Mamoru Hasegawa |
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Institution: | (1) Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., 3-6-6 Asahi-machi, 194 Machida-shi, Tokyo, Japan |
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Abstract: | We have compared the level of expression of several enhancer/promoters in human lymphoblastoid Namalwa KJM-1 cells when fused to a common reporter gene. A cassette containing the pro-urokinase (pro-UK) coding sequence followed by the rabbit -globin and simian virus 40 (SV40) 3 nontranslated region was used for evaluation of the enhancer activity. Cells containing Moloney murine leukemia virus (Mo-MuLV) promoter had an average of 10–20 fold higher expression levels of pro-UK than those containing other promoters, such as SV40 early gene promoter, human cytomegalovirus (hCMV) major immediate-early gene promoter, Rous sarcoma virus (RSV) promoter and chicken -actin gene promoter. The expression level of pro-UK under the control of Mo-MuLV promoter was 2–3 g/106 cells/day and was constant for more than 6 months. Furthermore, the production of a high producer clone, obtained by using dhfr gene coamplification, reached 30–40 g/106 cells/day. Thus, Mo-MuLV promoter showed the desired characteristics for efficient expression of foreign genes in Namalwa KJM-1 cells.Abbreviations dhfr
dihydrofolate reductase
- G-CSF
granulocyte colony-stimulating factor
- hCMV
human cytomegalovirus
- LTR
long terminal repeat
- Mo-MuLV
Moloney murine leukemia virus
- MTX
methotrexate
- pro-UK
pro-urokinase
- RSV
Rous sarcoma virus
- SV40
simian virus 40
- T3
triiodo-thyronine
- TRE
thyroid-hormone responsive element |
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Keywords: | efficient gene expression Moloney retroviral promoter Namalwa KJM-1 pro-urokinase |
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