determined to have an isoelectric point of 3.5 to 5 and will be analysed to ensure this result by use of an isoelectric focusing gel. This will establish whether the in process sample analyzed shows any potential for protein modification such as glycosylation or phosphorylation and following this run will be measured for the correct isoelectric point by use of 2D gel electrophoresis. (LifeTechnologies, 2014)
Thanks to TV shows like CSI, many people are familiar with the use of gel electrophoresis to separate macromolecules like DNA. However, gel electrophoresis can also be used to separate out proteins. Different proteins have different sizes, mainly due to the number of amino acid building blocks in their structure. Chemical modifications attached to the protein also affect its size. Different proteins also have different charges. This can result from both the types of amino acid used to construct
Electrophoresis is an analytical technique for the analysis of macromolecules like proteins and nucleic acids. This technique was discovered and first used in 1937 by a Swedish biochemist Arne Tiselius . The electrophoretic effect is based on the theory of Debye - Huckel - Onsager where this theory of electrolytic dissociation accept the fact that charged particles move up under the influence of electrostatic forces to an electrode of opposite charge is applied when a potential difference in a solution
chains” (Genetics Home Reference , 2014, p. xx-xx). “There are 20 different types of amino acids that can be combined to make a protein” (Genetics Home Reference, 2014, p. xx-xx). “The sequence of amino acids determines each protein’s unique 3-dimensional structure and its specific function” (Genetics Home Reference, 2014, p. xx-xx). “Proteins are complex molecules made up of carbon, hydrogen, oxygen and nitrogen (sometimes sulphur and phosphorus)” (TutorVista.com, 2014, p. xx-xx). There are four
The pK184hlyA and pBluescript plasmids were cut using EcoR1 and Pst1 restriction enzymes that cut at specific restriction sites. Gel electrophoresis was carried out to check if the digestion procedure was done successfully. Figure 2 shows the results of the electrophoresis. Lanes 5 and 7 indicate the fragments obtained when the plasmids are digested with both restriction enzymes, indicating the approximate fragment size for the hlyA gene, the pK184 plasmid and the pBluescript plasmid. This is useful
The purpose of the lab was to transform E.coli using the plasmid pRFP to promote the expression of antibiotic resistance as well as expression of the red fluorescent protein (RFP). The hypothesis was that if the transformation was successful, then the bacteria would express RFP because the arabinose would activate the plasmid’s red fluorescent protein, and show growth because pRFP allows E.coli to grow even in the presence of an antibiotic. The plasmid was combined with a sample of E.coli through
was spent dabbling in various activities, be it music, dance, sports, dramatics, quizzes or art, the one constant has always been my deep fascination with biology. This is what led me to choose biology and IT as my two high school specializations. The brief glimpse of what I got in my two years at high school left me in no doubt that Biotechnology Engineering was the field I wanted to pursue in my undergraduate studies. This helped me not only in exploring and understanding the research potential of
This causes the cells to take on an unusual “S” shape, therefore being named sickle-cell. The individual that expresses the disease must inherit two abnormal copies of the gene for haemoglobin from each parent, whereas carriers contain only one abnormal copy and do not show any symptoms. The defective gene is affected by a single base mutation of the β-globin gene, which replaces glutamic acid with