The intention of this study is bacterial transformation as the process of introducing the foreign DNA into the bacteria and gene expression as the process of obtaining information from the gene and using it for the synthesis of a functional gene product…
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This study looks into DNA as the blueprint of our life. Life without these molecules is not possible. DNA are transcribed into RNA and translated into proteins. These proteins are important for the biochemical functions of the cell. In bacteria, apart from DNA, there is extrachromosomal DNA called as plasmids. They are double stranded DNA which forms circles with size ranging from 1 kb to 200kb (kilobase). Plasmids are very advantageous for the genetic engineering. Plasmids code for many antibiotic regions and they have the ability to accept the gene of interest. The transformation of our gene of interest into the plasmid is called recombination and the bacteria are called recombinant bacteria. Thus plasmids can be used as cloning vehicles or vectors. These plasmids are not essential for the survival of bacteria, but in some instances, for survival in the different environments, they can provide some extra advantage. The best example is the survival of the bacterial cell in the presence of an antibiotic drug. Antibiotic resistant bacteria like Escherichia coli are used for the transformation of the gene of interest into the host cell. Transformation and cloning are the two important genetic engineering tools used for the expression of the foreign gene of interest in the given bacterial cell. Three conditions required for the transformation are: 1) the host into which the foreign DNA is inserted, 2) a method for the insertion of the DNA into the host cell, and 3) methods to identify the transformed cells and select them.
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This report examined the features of E coli in respect to the expression systems with an emphasis on the limitation number that have been addressed. For a high gene dosage to be achieved, the heterologous cDNAs are normally cloned into replicate plasmids in a fashion that is relaxed and are normally existence at 15-60 copies per cell.
ast tumour and normal tissue specimens, and compare the amplified target cDNA to a constitutively expressed house-keeping gene, glyceraldehyde-3-phosphate dehydrogenase (GAPDH).
Dysregulated cell cycle control is a primary mechanism of cancer (Deshpande, Sicinski & Hinds,
To confirm the success of transformation, the recombinant DNA was isolated and purified from the transformants followed by digestion using HIND III /EcoRV. The GFP protein was then purified from the transformants using hydrophobic interaction chromatography (HIC) and its purity confirmed using immunoblotting and SDS-Page.
It involves pasting the whole genome sequence of the organism given in a FASTA format then clicking the ORF finder that will automatically give the start and the stop codons alongside the longest pattern. In this case, the human genome was given and upon subjecting it to
The insert (16s rDNA = 1.5 kb; plasmid = 3.5 kb). The band on our slide is dark and broad therefore exact size determination is difficult. However, the size is slightly more than 4.0 kb, which is near the expected
EXPRESSION OF RECOMBINANT TICK HISTAMINE-BINDING PROTEIN TC11485 FROM Rhipicephalus microplus USING PPICZ-ALPHA IN Pichia pastoris Abstract Histamine-binding proteins (HBP) are ubiquitous biomolecules with pharmacologic importance, since they have an immunosuppressing activity, which can help prevent adverse health conditions such as histamine shock.
Moreover, the GFP (Green Fluorescent Protein) gene usually exists naturally in the form of specialized photogenic cells found in the jellyfish Aquorea Victoria umbrella. This fluorescent protein can also be expressed in the bacteria, Escherichia coli. When the fluorescent protein gets exposed to U.V light source of long wave band, it produces a green light.
Bacterial cell quantification can be done in several ways, but this experiment will be using a spectrophotometer to measure the relationship between the bacterial cell concentration and the absorbance of light. Results using known concentrations are often used to find the cell count of an unknown solution, and this is what will be achieved here.
4 Pages(1000 words)Lab Report
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