Plasmid

competent can still undergo transformation by being treated with various agents to artificially induce competence. Also, another factor that plays a role in transformation is plasmids. Plasmids are small circular pieces of DNA that assist with gene transfer in artificially induced transformation. In this lab, a previously made plasmid called pGLO was used to conduct the transformation process into Escherichia coli (E. coli). pGLO carries several genes such as β-lactamase, green fluorescent…

The bla gene codes for beta lactamase which promotes antibiotic resistance for drugs classified as being a part of the penicillin family. In the pGLO plasmid there is a unique gene regulation system which controls the transcription of the green fluorescent protein. In order for the GFP to be expressed, arabinose sugar must be present. Arabinose sugar reacts with the AraC protein, and this reaction initiates…

These particular strains are not pathogenic, however other strains can cause food poisoning if consumed. E. coli’s entire genome has been sequenced and the genes it codes for are relatively well known.1 E. coli DNA is in the form of a plasmid which is circular DNA. Plasmids are easy to manipulate, self-replicating, stable, and applicable to various organisms due to a high integration potential. E. coli research is often used to study gene expression and has a high potential of being applicable…

The first step in phage infection is the attachment of the phage to the host cell surface. This is typically accomplished by the recognition of a receptor on the outside of the bacterial cell wall such as an antigen, pilus or other structure. There is much variability from phage to phage in terms of which receptor they bind to. Bacteriophage can generally be classified into two categories, lysogenic and lytic (virulent) (Fig 6). The choice between the lytic and lysogenic cycle depends on the…

In this process, which is used mainly for short genomes such as bacteria, bacterial plasmids with different restriction enzymes are cut. The orientation of these restriction sites can be determined because each restriction enzyme cuts at a specific base sequence. This technique can be used to decipher an entire base sequence of the whole strand. In agriculture, it is important to locate the bacterial plasmids when introducing bacterial DNA into plant DNA. This is the technology that makes is…

DNA plasmid was added to each of the “+plasmid” and it expresses the ampicillin resistance gene. According to the data, that didn’t happen because the +pGLO LB/amp had 3 white colonies at the end of the bacterial transformation and the +pGLO LB/amp/ara had 2 colonies that glow at the end of bacterial transformation meaning that there was an error in our “+plasmid” which occurred during the lab. We did predict that the -pGLO LB/amp would remain the same with no colonies and -pGLO LB would grow…

How Ferritin was first isolated and how it is isolated now The protein ferritin is formed from two classes of subunits, H and L, in a ratio which vary in different cell types. The H-type are subunits associated with rapid uptake of iron and are predominantly from the red blood cells. Subunits that take up iron much slower are designated L-types. A third type subunit, M, is found in amphibians and is similar to the H subunit. The M-types are predominant in the liver. Once synthesized, ferritin…

Bacteria also known as prokaryotes are very ancient single-celled organisms; fossils show that they were widespread 1.5 billion years ago. Today bacteria make up the vast majority of prokaryotes on earth due to the unfavorable ability to adapt quickly to the surrounding environment. Bacteria have the capabilities to survive in areas of high heat, extreme cold, or even very acidic or alkaline conditions. This ability is used frequently to fend off the effects of antibiotics and antibacterial;…

BACTERIAL TRANSFORMATION By Derek Tang BIO 281 Lab section 80243 Abstract: The objective of this experiment was to observe the results of bacterial transformation in various growth conditions and determine how successful the altered organism is after the transformation. The organism used was E. Coli because singular cell organisms can have a new gene more easily inserted than multi-cellular organisms. The gene which was inserted is pGLO that carried the gene encoding GFP and a gene for…

Our goal within the plasmid isolation lab was to isolate the bacterial plasmid without any damage to the DNA. The formation of the clear lysate was essential because it allowed us to isolate the bacterial cells from removed media and later expose the DNA. Resuspension solution helped to resuspend the cells, after centrifugation, and prepare the cells for lysis solution. The lysis solution functions to degrade the cell membrane and spill its components. Then the Alkaline Protease functioned as…