Protein methods

Literature Review: Green Fluorescent Protein and Histidine Tagged Proteins Histidine and green fluorescent protein (GFP) tagging are important in molecular biology because they allow for purification, tracking, and quantification of target proteins (Ferrari et al., 2004; Cho et al., 2011; Deponte, 2012). Histidine tagging provides a method for isolating and increasing the amount of target protein recovered from a biological organism or mixed sample of proteins (Masek et al., 2011; Singh and…

Protein mutations are often introduced at the DNA level in the corresponding gene, which takes advantage of the central dogma of biology (DNA is transcribed to mRNA; mRNA is translated to proteins. The introduction of mutations in target genes has become trivial with the advances in synthetic biology. Major classes of library construction include (1) random mutagenesis, (2) recombination, (3) semi-rational design, and (4) scanning mutagenesis. Together, these classes of library construction…

natively folded proteins are not molecular weight dependent, the proteins would be traveling at different speeds in an electric field. In order to separate the proteins based on their molecular weight, the tertiary structure of the protein needs to be destroyed making the protein a linear molecule. Sodium dodecyl sulfate (SDS) plays an important role in this mechanism. Sodium dodecyl sulfate is an amphipathic detergent whose role is to break down the tertiary structure of a protein making it a…

Green Fluorescent protein and Green Fluorescent protein are suggested to be involved in bacterial transformation tool, which helps the scientists visualize the normal protein in cell. To test the hypothesis, the experiment had been collected the around 15 colonies from E.coli source plates. Those amounts of E.coli, then, were used to test under transformation of E.coli with Green and Blue Fluorescent Proteins procedure. As the result, the E.coli that had been injected pFluoroGreen and…

for 30 min at room temperature. Then, trypsin/chymotrypsin- digested samples of KPNB1 obtained from the UV-crosslinking experiments with various concentrations of compound 1 were analyzed by nanoflow liquid chromatography and tandem mass spectrometry. The tandem mass spectra of the trypsin/chymotrypsin-digested KPNB1-compound 1 adducts showed 10 peptides coupled to compound 1 (Table S1, the Supplementary Information). Among these adducts, we found that the amounts of three of the products,…

(2007) states that “Eggs are used for other purposes besides food. For example, they are used in vaccine preparation and for other laboratory purposes.” (pg. 118). Eggs are used in many bakeries since they are a great base in recipes. The egg’s protein becomes solid and can expand when heated, making things like cakes fluffy. Making use of the entire egg in a recipe gives the product moisture and a firm texture. In addition, eggs are a main part of what keeps ice crystals from forming in ice…

amplification method in molecular biology, it was invented by Kary Mullis while working in Emeryville, California for Cetus Corporation, one of the first biotechnology companies. His invention won him a Nobel Prize in Chemistry in 1993. PCR has revolutionized the field of molecular biology as it has enabled researchers to perform experiments easily that previously had been unthinkable. Before the mid-1980s, when PCR was developed, molecular biologists had to use laborious and time-consuming…

Transforming Genetic Material To E. Coli Using Plasmids Proteins are created within the body through a process known as central dogma. This process is the transcription of DNA to RNA and the translation of RNA to proteins. The type of protein created relies on what type of messenger RNA or mRNA that was produced during the transcription of DNA. In our experiment, we looked at the specific LacZ gene that codes for beta-galactosidase, which is a protein/enzyme that can break down lactose into its…

Introduction: One of the most common mass spectrometry–based proteomic methodologies used to day is the bottom up protein identification. First, the protein has to be digested into peptides using a protease. Then, the peptide will be submitted to the URMC Proteomics Resource and Laboratory for analysis using LC-MS/MS tandem mass spectrometry. In this approach, the peptides will be separated and introduced into a mass spectrometer using reversed phase liquid chromatography (RC-LC). This…

sieve; Wwheat in 75μm sieve = weight of wheat kernels in 75 189 μm sieve; Wpan = 190 weight of empty pan; and Wmaize meal = weight of maize flour. 191 Determination of Crude Protein 192 The crude protein content of the roasted maize was determined in duplicate by Dumas combustion 193 method as described in AACCI Approved Method 46–30.01 (AACC, 1999) using a Leco, model 194 FP-528 (Leco Africa, Kempton Park, South Africa). Calibration of the instrument was done using 195 an alfalfa standard with…