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Your assignment is to prepare and submit a paper on the production and isolation of green fluorescent protein.
Your assignment is to prepare and submit a paper on the production and isolation of green fluorescent protein. The study was carried out to explore the production and isolation of Green Fluorescent Proteins. The study of GFP was particularly of importance based on its use in biotechnology during the creation of genetically modified organisms. GFP is one of the notable reporter genes found in pGLO, normally used as a vector aiding in the production of GMOs. In which case, pGLO is a placid containing three genes: Bla that aids in the transformation of bacteria resistance, an area that controls the expression of GFP and GFP, per se, exists as a fluorescent protein, which gives the characteristic green glowing color. The phenomenal green color of GFP acted as a core material in the production of Alba, the green fluorescent rabbit.
GFP is a major compound confirmed to generate fluorescence for mammals, especially marine organisms. This confirms the wide routine use of GFP in various researches. In which case, a given gene is cloned in the presence of GFP then expressed to allow it to link well with the latter (GFP). From this, the protein will start glowing green thereby allowing for easy tracking of its location among other cells in an organism. The presence of AmpR, the ampicillin resistance gene allows for selection. The expression of GFP in this plasmid serves to evaluate whether the cells have gone a complete transformation.
The specific purpose of the study is to explore the production and isolation of Green Fluorescent Protein (GFP) (Chess, 2009). This process will include several related laboratory techniques, including bacterial transformation, selection of transformed cells, Growth of recombinant bacteria and isolation of the GFP. The lab presents the results obtained for each laboratory technique and compares them with the already existing knowledge.
Methods
Two test tubes were obtained and labeled +pGLO and another –pGLO. 250 µl of transformation solution (CaC12) were transferred using a sterile transfer pipette to the test tubes. All the tubes were placed on ice. Single colony bacteria from the starter were obtained from the starter plate followed by picking up the +pGLO tube and immerse the loop into the transformation solution at the bottom of the tube. The UV lamp provided was used to examine the pGLO plasmid DNA solution.
