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Why is ampicillin added to the pound medium?
Ampicillin is an antibiotic that is usually used as a reporter gene in cloning. A plasmid containing the ampicillin resistance gene (as well as another target gene within the plasmid) is introduced into the bacterial host. If the bacterium has taken up the plasmid and is expressing the plasmid, it will be resistant to ampicillin. LB is used as a growth medium and ampicillin to verify the plasmid is within the bactrium. No growth means no plasmid in the bacterial host...
What enzyme is produced by cells transformed with plasmid lux that is not produced by cells transformed with pUC18?
The enzyme produced by cells transformed with plasmid lux that is not produced by cells transformed with pUC18 is luciferase. This enzyme is responsible for the bioluminescent properties of animals like fireflies and glowworms. Cells transformed with plasmid lux will emit light in the presence of the substrate luciferin, whereas cells transformed with pUC18 will not.
A population of cells carrying a desired plasmid is called a?
A population of cells carrying a desired plasmid is called a transformed population.
What is the ampr gene?
The ampr gene encodes for the enzyme beta-lactamase, which confers resistance to ampicillin in bacteria. This gene is often used as a selectable marker in molecular biology experiments to identify transformed cells that have taken up a plasmid with the gene.
Why GFP colonies are observed in the plate with pQE30 and GFP construct?
Go to http://www.tsienlab.ucsd.edu/Images.htm you will find bacterial colonies will and can express functional fluorescent proteins. As for the plasmid pQE30, it looks to fit into the category of expression at this level.
Why is ampicillin added to the pound medium?
Ampicillin is an antibiotic that is usually used as a reporter gene in cloning. A plasmid containing the ampicillin resistance gene (as well as another target gene within the plasmid) is introduced into the bacterial host. If the bacterium has taken up the plasmid and is expressing the plasmid, it will be resistant to ampicillin. LB is used as a growth medium and ampicillin to verify the plasmid is within the bactrium. No growth means no plasmid in the bacterial host...
What enzyme is produced by cells transformed with plasmid lux that is not produced by cells transformed with pUC18?
The enzyme produced by cells transformed with plasmid lux that is not produced by cells transformed with pUC18 is luciferase. This enzyme is responsible for the bioluminescent properties of animals like fireflies and glowworms. Cells transformed with plasmid lux will emit light in the presence of the substrate luciferin, whereas cells transformed with pUC18 will not.
A population of cells carrying a desired plasmid is called a?
A population of cells carrying a desired plasmid is called a transformed population.
What is the ampr gene?
The ampr gene encodes for the enzyme beta-lactamase, which confers resistance to ampicillin in bacteria. This gene is often used as a selectable marker in molecular biology experiments to identify transformed cells that have taken up a plasmid with the gene.
Why GFP colonies are observed in the plate with pQE30 and GFP construct?
Go to http://www.tsienlab.ucsd.edu/Images.htm you will find bacterial colonies will and can express functional fluorescent proteins. As for the plasmid pQE30, it looks to fit into the category of expression at this level.
What is the purpose of the ampicillin in the transformation procedure?
The ampicillin resistance gene (AmpR) is called a selectable marker gene and is incorporated into several plasmids that are commonly used in a wide range of genetic engineering and molecular biology experiments. The function of a selectable marker gene is to provide the host containing the plasmid a certain property that is not inherently present in the host. For example, the AmpR gene codes for a protein that makes cells containing this gene resistant to the antibiotic ampicillin. Therefore, when plasmids are successfully transfected into bacterilal cultures, one can assess which colonies have taken up the plasmid by looking at which ones survive onampicillin-containing media. Those that do not survive do not have the plasmid. The surviving colonies can then be picked up and carried on to the next stage of experimental research.
How does pBR322 work as cloning vector?
pBR322 is a plasmid vector that contains an origin of replication for replication in E. coli, as well as antibiotic resistance genes for ampicillin and tetracycline. It also has unique restriction sites for easy insertion of foreign DNA. Once the foreign DNA is inserted into the vector, the plasmid can be transformed into E. coli cells where it replicates and expresses the inserted DNA.
What is meant by a control plate in pglo experiments?
a control plate in your particular case is a ampicillin plate with no inoculation of your transformed bacteria. its purpose to make sure that during the process of handling the plate there were no contamination by bacteria in the surrounding.
Suppose you transform bacteria with a recombinant DNA plasmid and by mistake grow the transformed cells without an antibiotic . What result would you observe?
If you grow transformed bacteria containing a recombinant DNA plasmid without an antibiotic, you would likely observe that only a small proportion of the cells that successfully took up the plasmid will survive, while the majority of non-transformed cells will also grow. However, the transformed cells may not express the gene of interest or provide any selective advantage, resulting in no significant difference in growth compared to the non-transformed cells. Over time, the population would likely consist mostly of non-transformed cells, as they do not require any selective pressure to thrive.
Bacterial cells which have been transformed with a plasmid are allowed to grow so that they the plasmids everytime they divide?
The transformed bacterial cells will replicate the plasmid along with their own genomic DNA each time they divide. This allows for amplification of the plasmid within the bacterial population. The plasmid can carry genes for antibiotic resistance, gene expression, or other functions that can be advantageous for the bacteria in certain conditions.
Is it true cells containing a plasmid with an antibiotic resistance gene will die in the presence of the antibiotic?
false
How will you identify cells carrying the recombinant plasmid?
The transformants are selected for on agar containing an appropriate antibiotic. For example if your recombinant plasmid contains a kanamycin cassette, then only the cells containing the plasmid will grow on an agar plate containing kanamycin. PCR can then be performed on the colonies to ensure they contain your gene of interest on the plasmid.
