+ Resolve Article
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter

+ Translate
+ Subscribe to Site Feed
GeoScience Most Shared ContentMost Shared Content

Expression and analysis of green fluorescent proteins in human embryonic kidney cells by capillary electrophoresis

, : Expression and analysis of green fluorescent proteins in human embryonic kidney cells by capillary electrophoresis. Analytical Biochemistry 268(2): 262-269

The green fluorescent protein (GFP) has attracted much interest as a reporter for gene expression. In this paper, application of capillary electrophoresis with laser-induced fluorescent (CE-LIF) for quantitation of green fluorescence protein in cellular extracts and single cells is investigated. The S65T mutant form of GFP protein was successfully expressed in human embryonic kidney (HEK293) cells, and its production was confirmed by fluorescence microscopy and CE-LIF. The mass limit of detection for the mutant S65T was 5.3 x 10(-20) mol, which was better than that for the wild-type GFP by a factor of six. Detection of a small amount of GFP is difficult by conventional techniques such as fluorescent microscopy due to interference from cell autofluorescence at low GFP concentrations. The HEK293 cells were transfected with the GFP plasmid that produced S65T-GFP. Transient production of S65T protein was detected 2 h after the transfection and reached a maximum after 48 h. The protein concentration began to decrease significantly after 96 h. Single cell analysis of HEK293 cells after transfection with GFP plasmid indicate a nonuniform production of S65T-GFP protein among cells.


PMID: 10075816

DOI: 10.1006/abio.1998.2975

Other references

Anonymous, 1998: Culture of rhubarb. The culture of rhubarb (Rheum spp.) is described including notes on propagation, planting, cultivars, manuring, forcing, the control of pests and diseases, lifting and harvesting, with estimates of costs and returns.

Béguin, S.; Keularts, I.; A.D.eri, R.; Bellucci, S.; Caen, J.; Hemker, H.C., 2004: Fibrin polymerization is crucial for thrombin generation in platelet-rich plasma in a VWF-GPIb-dependent process, defective in Bernard-Soulier syndrome. Defective prothrombin consumption has been reported in the proband case of Bernard-Soulier syndrome (BSS). There is no consensus, however, on whether the formation of platelet procoagulant activity (PPA) is impaired in BSS and, if so, whether this...

Boiardi, A.; Bussone, G.; Negri, S., 1977: Critical assessment of the local curare test in myasthenia gravis. In a group of 60 patients of which 30 were affected by myasthenia, we used the regional curare test. The results were evaluated from a critical point of view and were then compared with the results obtained by the repetitive stimulation of the cir...

Bekric, V.; Bozovic, I.; Pavlovski, Z.; Masic, B., 1990: Effects of differently treated soya-bean as a component in broiler feeds. Meal was prepared from raw soyabeans or soyabeans which had been roasted with continual mixing, cooked in a pressure cooker or cooked in a microwave oven. For 56 days boiler chickens were given diets prepared with the 4 different meals and a fifth...

Gueyffier, F.; Boutitie, F.; Boissel, J.P.; Pocock, S.; Coope, J.; Cutler, J.; Ekbom, T.; Fagard, R.; Friedman, L.; Perry, M.; Prineas, R.; Schron, E., 1997: Effect of antihypertensive drug treatment on cardiovascular outcomes in women and men. A meta-analysis of individual patient data from randomized, controlled trials. The INDANA Investigators. Trials of drug therapy for hypertension have shown that such therapy has a clear overall benefit in preventing cardiovascular disease. Although these trials have included slightly more women than men, it is still not clear whether treatment benefi...

Michalowski, J.; Halaburda, P., 2001: Flow-injection chemiluminescence determination of epinephrine in pharmaceutical preparations using raw apple juice as enzyme source. Raw apple juice exhibits the activity of polyphenol oxidase which can be employed for the determination of mono-, di- and polyhydric phenols. The chemiluminescence detection in the two-channel manifold was used to develop the new procedure for det...

Daniel, M.K., 1972: The Hammond collection of reprints on milk. Author catalogue. Subject catalogue. These 2 volumes are catalogues of the collection of about 2400 reprints dating from 1899 covering all aspects of milk production as well as milk and milk products.

Spielmann, H.; Seiler, A.; Bremer, S.; Hareng, L.; Hartung, T.; Ahr, H.; Faustman, E.; Haas, U.; Moffat, G.J.; Nau, H.; Vanparys, P.; Piersma, A.; Sintes, J.Riego.; Stuart, J., 2006: The practical application of three validated in vitro embryotoxicity tests. The report and recommendations of an ECVAM/ZEBET workshop (ECVAM workshop 57). The major limitations of the validated in vitro embryotoxicity tests identified at the 2003 workshop were immediately taken into account by ECVAM in a proposal for an Integrated Project in the EU 6th Framework Programme, entitled ReProTect (22). S...

Saines, J.C., 1997: Phenomenon of sudden death: Part I. Accident and Emergency Nursing 5(3): 164-171

Bang, N.U.; Iversen, K.; Jagt, T.; Tobiassen, G., 1959: Serum glutamic-oxaloacetic transaminase activity as an index of centrilobular liver cell necrosis in cardiac and circulatory failure. Acta Medica Scandinavica 164: 385-393