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What type of gene is used to distinguish bacteria that carry a plasmid containing foreign DNA from those that don' t?
Wiki User
∙ 14y ago
The plasmid that contains foreign DNA is engineered to also carry an antibiotic resistance gene. This antibiotic resistance gene codes for a protein that is able to inactivate an antibiotic thus keeping the cell alive.
In the absence of the antibiotic resistance gene, the cells would not survive when exposed to an antibiotic.
After transfection (the process of inserting the plasmid carrying the foreign gene into cells), the cells are gown in media containing an antibiotic.
Cells that contain the plasmid (and therefore contain the antibiotic resistance gene) are able to survive in this medium. Cells that do not contain the plasmid (and therefore lack the antibiotic resistance gene) do not survive in this medium.
The process described above is called selection
Wiki User
∙ 14y ago
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Which organelles inside bacteria make green fluorescent protein?
Organelles inside bacteria do not 'make' GFP. GFP can however be expressed in bacteria by adding the coding sequence for GFP to their genome. This can be done by using a plasmid virus containing the GFP gene sequence. When combined with the plasmid, a very small number of bacteria will take up the plasmid's genome into their own. These bacteria can be isolated by using the 'transform or die' mechanism. Bacteria that do not contain the plasmid are preferentially killed by adding an antibody to the agar. The normal bacteria will not be resistant to this, and will die, but the bacteria which have taken up the plasmid into their genome will also contain an antibody resistance gene. This allows them to live in the presence of the antibody. Since GFP was also added to these bacteria's genome, they will fluoresce.
What organisms have plasmid?
Bacteria
What is col plasmid?
A plasmid (free bit of DNA) in bacteria that produces colicin to kill other strains of bacteria.
A plasmid a foreign gene and recombinant DNA would be associated with what type of organism?
A plasmid is a double stranded cirucular DNA, used as a vector in cloning. A gene of interest can be ligated into the this to form a chimeric DNA or rDNA. This can be transformed to a bacteria for propagation of the clones (you can amplify them by these transformed bacteria).
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.
A gene that makes it possible to distinguish bacteria that carry the plasmid from those that dont?
Genetic marker.
How is it possible to splice foreign DNA into a plasmid without altering the bacteria's ability to function normally?
A plasmid is an extra-chromosomal element. Strictly speaking, it is not a part of the bacterial mail genome. Plasmids, when first discovered in bacteria, were observed to contain genes that confer special features to the bacteria (like drug resistance). Also, a plasmid is capable of autonomous replication, which means it has its own origin of replication and does not have to wait for the bacterial genome to be replicated. When a foreign gene is inserted into a plasmid, the plasmid has to be transformed into the bacteria. Once transformed, the cDNA in the plasmid is able to replicate every time the plasmid replicated. This does not have any affect on the normal functioning of the bacteria. The organism is merely being used to make copes of the foreign gene and/or harvest an important protein product.
Bacteria containing a plasmid into which the eukaryotic gene has integrated would grow where?
the ampicillin broth and the nutrient broth
Bacteria can take up foreign DNA molecules packaged in plasmids?
yes. Some bacteria can take up plasmids present in the solution around them or by getting the plasmid from another bacteria which possess a special "fertility" F-plasmid.
What one plate would you first inspect to conclude that the transformation occurred successfully Why?
If you transform bacteria with a plasmid containing a selection marker (such as an antibiotic resistance gene) and plate the transformed bacteria on a plate suited for selecting for plasmid-containing bacteria (such as a plate containing an antibiotic that only those bacteria with antibiotic resistance can survive), then simply inspecting whether colonies are present on the plate will suffice in determining whether the transformation succeeded. If no colonies are found, that means no bacteria got the antibiotic resistance gene on the plasmid and the transformation was unsuccessful. If some colonies are found, that means some bacteria contain the plamis containing the antibiotic resistance gene and those colonies can the transformation was successful.
Describe the use of plasmids as vectors in biotechnology?
Plasmids are often used as expression vectors in biotechnology. Plasmids are small, circular or linear pieces of DNA containing non-essential genes that are found in all life, although much more common in prokaryotes, especially bacteria. These genes confer abilities such as metabolizing a previously unusable compound, building an amino acid previously unbuildable, or even antibiotic resistance. Plasmids are used in research to induce the expression of a gene usually not found in the given organism. For example, you can construct a plasmid with a bacterial promoter connected to the gene for a human protein through a process called 'cloning'. The plasmid with the human gene can then be introduced into bacteria by transforming a competent gram-negative with the plasmid. Usually the plasmid also has an antibiotic resistance gene in addition to the target gene. This antibiotic resistance can be used to select for bacteria containing the plasmid. For example, the most common resistance gene is ampicillin resistance gene. If you grow the transformed bacteria in a culture containing ampicillin, only bacteria containing the antibiotic resistance, and therefore containing the plasmid, can survive. This will ensure that what you have is a pure culture of bacteria containing the plasmid. After selection, these bacteria can be cultured in suitable media to increase their numbers to a point that their production of the human protein becomes appreciable. Then these bacteria are usually lysed (killed) to extract the protein. Sometimes, however, these bacteria can also be made to secrete the protein into the medium.
Which organelles inside bacteria make green fluorescent protein?
Organelles inside bacteria do not 'make' GFP. GFP can however be expressed in bacteria by adding the coding sequence for GFP to their genome. This can be done by using a plasmid virus containing the GFP gene sequence. When combined with the plasmid, a very small number of bacteria will take up the plasmid's genome into their own. These bacteria can be isolated by using the 'transform or die' mechanism. Bacteria that do not contain the plasmid are preferentially killed by adding an antibody to the agar. The normal bacteria will not be resistant to this, and will die, but the bacteria which have taken up the plasmid into their genome will also contain an antibody resistance gene. This allows them to live in the presence of the antibody. Since GFP was also added to these bacteria's genome, they will fluoresce.
What organisms have plasmid?
Bacteria
What is col plasmid?
A plasmid (free bit of DNA) in bacteria that produces colicin to kill other strains of bacteria.
Why does bacteria that contains plasmid glow in uv light?
the plasmid contains a certain gene, which codes for the "Green Flourescent Protein." So you put the plasmid in the bacteria, the plasmid starts making that protein in the bacteria, and boom you've got glowing bacteria. works for bunnies and monkeys too, apparently =)
Compare the transformation of a bacterium cell with the transformation of a plant cell?
In bacteria, if the plasmid containing the foreign DNA manages to get inside a bacterial cell, this sequence ensures that it will be replicated. In Plant Cells, if transformation is successful the recombinant DNA is integrated into one of the chromosomes of the cell.
A plasmid a foreign gene and recombinant DNA would be associated with what type of organism?
A plasmid is a double stranded cirucular DNA, used as a vector in cloning. A gene of interest can be ligated into the this to form a chimeric DNA or rDNA. This can be transformed to a bacteria for propagation of the clones (you can amplify them by these transformed bacteria).
