Biology: Scientific Representation
Biology: Scientific Representation: Expression and purification of His-GFP
Figure 1 depicts the results from the expression and purification of histamine-green fluorescent protein (GFP), a molecule that is used as a molecular tag because it can be inserted into genes. Zeyton (et al., 2003) explained that GFPs are good for studying variant genes because of their “intrinsic fluorescence, high stability, expression and solubility” (p. 1473). The GFP can glow with green fluorescence under ultra-violet making it useful for identifying the reactions of variant genes with bacteria. Zeyton (et al., 2003) called them ‘”fluorobodies’ (which) have been used effectively in enzyme-linked immune-sorbent assays (ELISAs)” (p. 1473). In Figure 1 the right hand side letters naming the rows symbolize different dilutions; the last one which is labeled the PBS is the protein background signal. A control is used (such as with an antigen) to compare to molecules without an antigen (see right bottom left). Water baths at different temperatures are used for the appropriate time needed in the experiment (bottom labels). The labels at the top are (+) meaning an antigen is present and (-) meaning an antigen is not present.
Figure 1. Expression and purification of His-GFP, Optical Density of Sample
N: Negative Control is washing with no his just with GFP
C: Crude Fraction with protein which is the his-GFP tag
F: Flow through fraction Wash and Purification are done at the same time
E: Eluate (Buffer) Washing with the buffer
Reference
Zeytun, A., Jeromin, A., Scalettar, B., Waldo, G., & Bradbury, A. (2003). Fluorobodies combine GFP fluorescence with the binding characteristics of antibodies. Nature Biotechnology, 21(12), 1473-1479.
