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Viruses
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TaigaA recombinant plasmid can be introduced into a bacterium through a process called transformation, where the bacterium is made competent to take up the plasmid DNA. This can be achieved using methods such as heat shock, electroporation, or chemical treatment. Once inside the bacterium, the plasmid can replicate independently and express the desired gene.
There are two main methods: using chemicals and using electricity.
A combination of certain solutions, such as calcium chloride, and exposure to particular temperatures, makes bacteria more competent (i.e. "receptive", if you like).
Another way to allow plasmids to enter bacteria is to apply a pulse of electricity; this method is known as electroporation. It is more efficient and consistent, but more expensive.
These methods are the result of trial-and-error (officially called empiricism), and just why they work nobody knows.
The recombinant DNA can be inserted into a bacterial cell by a method called transformation/calcium chloride treatment. It is the free uptake of DNA by the competent cell. In this method the recombinant DNA is added to the broth culture containing bateria, to this CaCl2 is added and incubated for 12 hrs. The recombinant DNA gets precipitated on the host cell due to CaCl2. This is subjected to heat shock treatment ( 42 degree celsius ) for 2 minutes during which the pores are created on the host cell wall and the recombinant DNA gets integrated with the host cell DNA.
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What occurs first in the production of a recombinant plasmid?
In the production of a recombinant plasmid, the DNA of interest (insert) and the plasmid vector are both cut with restriction enzymes to create compatible ends. These cut fragments are then ligated together using DNA ligase to produce the recombinant plasmid.
What is recombinant Plasmid?
A plasmid is a small, circular, double-stranded DNA molecule that is distinct from a cell's chromosomal DNA. ... Researchers can insert DNA fragments or genes into a plasmid vector, creating a so-called recombinant plasmid. This plasmid can be introduced into a bacterium by way of the process called transformation.
What is the term for a plasmid that contains a foreign gene?
Recombiant DNA
When is a plasmid considered a recombinant plasmid?
A plasmid is considered recombinant when it contains DNA sequences from two different sources that have been artificially combined, often through genetic engineering techniques like restriction enzyme digestion and ligation. This results in a plasmid with modified or additional genetic material compared to its original form.
What is the most logical sequence of steps for splicing foreign DNA into a plasmid and inserting the plasmis into a bacterium?
Cut the plasmid and foreign DNA with the same restriction enzyme to create complementary sticky ends. Mix the cut plasmid and foreign DNA together and ligate them using DNA ligase. Introduce the ligated plasmid into the bacterium using a method like transformation, where the bacterium uptakes the plasmid. Select for transformed bacteria using antibiotic resistance or another selectable marker on the plasmid.
