geoscience.net logo
+ 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

Dual-function receptor protein for analysis of gene expression in living cells


, : Dual-function receptor protein for analysis of gene expression in living cells. Biotechniques 25(5): 848-856, Nov

The firefly luciferase (Luc) protein and the jellyfish green fluorescent protein (GFP) are two commonly used molecular reporters that can be detected noninvasively in living cells. The properties that make GFP or Luc useful for a particular experimental application are quite distinct. A recombinant protein with both fluorescent and bioluminescent characteristics might take advantage of the strengths of both reporters. An expression vector encoding a chimeric protein in which GFP was tethered to Luc through a 19-amino acid linker was prepared and characterized Western blotting with antibodies specific for either GFP or Luc showed that a protein of appropriate size was expressed in transfected cells. Fluorescence microscopy revealed bright green fluorescence from transfected cells, indicating proper formation of the GFP chromophore. Luc enzymatic activity in protein extracts from transfected cells showed that Luc was fully functional. The treatment of living cell cultures stably expressing the GFP-Luc fusion protein with the protein translation-inhibitor cycloheximide (Chx) was used to show that the half-life for Luc protein activily was approximately 2 h at 37degreeC. The utility of this dual-function reporter protein was shown by the identification of single living cells expressing the chimeric protein within a population by fluorescence microscopy, followed by quantification of Luc activity from the same living cells.

US$29.90


Other references

Missmahl, H.P.; Strohmaier, B., 1964: Deposition of basic substance on collagen fibers. Quantitative evaluation of the phenol reaction of von Ebner was carried out with regard to the metachromasia at the collagen fiber of the corium and the subcutaneous tissue. Treatment with hyaluronidase and incubation with heparin or chondroitin-s...

Kondrat ev S.I. (Kondrat yev S.I.; Romanov A.S.; Vnukov Y.L., 2007: Distribution of hydrochemical characteristics within continental slope in the northwestern Black Sea. Peculiarities of spatial distribution of hydrochemical characteristics in the anti-cyclonic eddy in the northwestern Black Sea in May, 2004 are investigated. Surprisingly high location of 3 mu M isosulfide at the isopycn sigma (sub 1) = 15.88 kg/m...

Hofling-Lima, A.Luisa.; Branco, B.Castelo.; Romano, Aé.C.; Campos, M.Q.S.; Moreira, H.; Miranda, D.; Kwitko, S.; de Freitas, D.; Casanova, F.H.; Casanova, F.; Sartori, M.; Schor, P.; Souza, L.B., 2004: Corneal infections after implantation of intracorneal ring segments. Purpose: To report risk factors, clinical course, and outcome in patients with infectious keratitis following implantation of intracorneal ring segments (ICRS). Methods: The records of 8 patients with culture-proven infectious keratitis after ICRS...

Zatorre, R.J.; Gandour, J.T., 2007: Neural specializations for speech and pitch: moving beyond the dichotomies. The idea that speech processing relies on unique, encapsulated, domain-specific mechanisms has been around for some time. Another well-known idea, often espoused as being in opposition to the first proposal, is that processing of speech sounds ent...

Corradi, L.S.; Góes, R.M.; Vilamaior, P.S.L.; Taboga, São.R., 2010: Increased androgen receptor and remodeling in the prostatic stroma after the inhibition of 5-alpha reductase and aromatase in gerbil ventral prostate. Prostate require high levels of steroidogenic enzymes such as 5alpha-reductase (5alpha-r) and Aromatase (Aro) for the formation of active steroids. Dihydrotestosterone (DHT), the prostate dominant androgen, is converted from testosterone (T) by th...

Fiorucci, G., 2009: On the meaning of the erythrocyte sedimentation rate. American Journal of Clinical Pathology 132(5): 806-806

Flourie, B.; Nancey, S., 2007: Functional properties of probiotics. The ability to resist gastric acidity, bile salts, and the intestinal environment varies widely among probiotics. The strains found in yoghurt are destroyed in the upper digestive tract and only 1% is delivered to the ileum. In comparison, many st...

Loomis, John B., 2002: Integrated public lands management: principles and applications to national forests, parks, wildlife refuges, and BLM lands. 2nd edition. Unknown

Shinohara, Y.; Kumai, T.; Higashiyama, I.; Hayashi, K.; Matsuda, T.; Tanaka, Y.; Takakura, Y., 2016: Histological and molecular characterization of the femoral attachment of the human ligamentum capitis femoris. The ligamentum capitis femoris (LCF) has increased in clinical significance through the development of hip arthroscopy. The histological pathologies and molecular composition of the femoral attachment of the LCF and the degeneration caused by LCF...

Zachrisson, B., 1971: Toothbrushing in patients with orthodontic appliances. Den Norske Tannlaegeforenings Tidende 81(9): 686-693