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One of the most common ways these days is from cDNA. RNA is extracted from human cells, purified, and then treated with an enzyme called reverse transcriptase which is able to make DNA from RNA templates (this DNA made from RNA is called cDNA). The advantage of using cDNA is that in the genome human genes are typically distributed across multiple exons spread over tens or even hundreds of thousands of basepairs of DNA. Such a massive segment of DNA is extremely hard to manipulate and far too large to insert into a plasmid. However, in cDNA, all the introns have been spliced out (because cDNA is made from mature mRNA).
To isolate a particular gene from cDNA, PCR is often used to selectively amplify one gene's cDNA using specific primers. To insert the amplified cDNA into a plasmid, the traditional approach was to use restriction enzymes - enzymes that cut precise DNA sequences. The great thing about many restriction enzymes is that they cut DNA but leave behind "sticky ends". Thus if you cut both your cDNA and a plasmid with a particular restriction enzyme, the resulting sticky ends will allow the human cDNA to be taken up by the plasmid (the sticky ends will mesh). The sticky ends will have to be sealed by an enzyme called DNA ligase.
However, there are other ways these days - often involving recombination to insert the PCR product directly into a plasmid without resorting to restriction enzymes and ligations.
What else can I help you with?
In the process of human gene cloning using recombinant plasmids what is the bacterial plasmid?
The bacterial plasmid is a small circular DNA molecule that is used as a vector to carry the gene of interest in gene cloning experiments. It is introduced into bacteria, where it replicates independently from the bacterial chromosome. The gene of interest is inserted into the plasmid using restriction enzymes and ligase.
How can bacteria be transformed with recombinant plasmid?
Bacteria can be transformed with recombinant plasmid by introducing the plasmid into the bacterial cell through a process called transformation. This allows the bacteria to take up the recombinant DNA from the plasmid and express the desired gene or trait encoded in the DNA.
How can one effectively clone a gene into a plasmid?
To effectively clone a gene into a plasmid, the gene of interest and the plasmid are cut with the same restriction enzymes to create compatible ends. The gene is then inserted into the plasmid using DNA ligase to seal the ends. The plasmid is then introduced into a host cell, such as bacteria, where it can replicate and express the cloned gene.
How can one effectively insert a gene into a plasmid?
To effectively insert a gene into a plasmid, one can use restriction enzymes to cut both the gene and the plasmid at specific sites. The cut gene can then be inserted into the plasmid, and DNA ligase can be used to seal the pieces together. This process is known as molecular cloning.
The takes up the plasmid. It now contains the human gene?
I think I know the answer... it's 5
In the process of human gene cloning using recombinant plasmids what is the bacterial plasmid?
The bacterial plasmid is a small circular DNA molecule that is used as a vector to carry the gene of interest in gene cloning experiments. It is introduced into bacteria, where it replicates independently from the bacterial chromosome. The gene of interest is inserted into the plasmid using restriction enzymes and ligase.
How does a human insulin genes become part of a plasmid?
1. Scientists remove plasmids, small rings of DNA, from bacterial cells. 2. An enzyme cuts open the plasmid DNA. The same enzyme removes the human insulin gene from its chromosome. 3. The human insulin gene attaches the open ends of the plasmid to form a closed ring. 4. Some bacterial cells take up the plasmids that have the insulin gene. 5. When cells reproduce, the news cells will contain copies of the engineered plasmid. The foreign gene directs the cell to produce human insulin.
How can large quantities of protein be produced from a bacterial colony containing the gene of interest?
Large quantities of protein can be produced by expressing the gene of interest in a bacterial colony such as E. coli. This is typically achieved by cloning the gene into a plasmid, transforming the plasmid into the bacterial cells, and inducing protein expression. The bacterial colony can then be grown in a culture medium optimized for protein production to maximize yields.
Can genetic engineering techniques treat cystic fibrosis?
Yes, there are two similar techniques in which i am aware of.AdenovirusesFirstly adenoviruses are made harmless by interefering with a gene involved in replication. A healthy from of the CFTR gene is extracted and cut with restriction endonucleases, the same enzyme is used to cut a bacterial plasmid. The gene and plasmid are mixed together along with DNA ligase to anneal the phosphosugar framework of the DNA fragment and bacterial plasmid. The plasmid is then mixed with epithelial cells. The plasmid is then isolated and purified and places into adenoviruses. These are then placed onto the nostrils of individuals with cystic fibrosis. The viruses find their way to epithelial cells in the airways and injected their DNA. The DNA contains the functional CFTR gene, the cells can then produce fucntional CFTR proteins.LiposomesA healthy gene is extracted from a human. This gene is then inserted into a bacterial plasmid, in a similar manner as discussed above. The bacterial plasmids are then inserted into bacteria. These are allowed to grow and divide, producing large quantities of the plasmid, with the required gene. These plasmids are then extracted and coated in a lipid soluble substance. They are then put into nasal sprays and sprayed onto the nostrils of effected individuals.
What does the bacterial cell reproduce that the human gene codes for?
the bacterial cell reproduces the bacterial chromosome that the human gene codes for.
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...
A plasmid that contains a gene for human growth hormone is and example of what?
A plasmid containing a gene for human growth hormone can be used in genetic engineering to produce recombinant human growth hormone. This plasmid can be introduced into host cells, such as bacteria, for the production of the hormone on a large scale.
What is one way to determine whether a bacterial culture has received a recombinant plasmid?
The plasmid have a "reporter gene" inside it, generally resistance to specific antibiotic. the plasmid is transformed into bacteria that don't have resistance to that specific antibiotic drug, and than the cultured on a petri-dish that contain the antibiotic drug. Only bacteria that had receive the plasmid will have resistance and grow, all the other will die.
What is a multicopy plasmid?
Every plasmid has a copy number that reflects the average number of copies of a certain plasmid inside a host cell(usually a bacterial cell). So a multicopy plasmid, exist in multiple copies in any given bacteria. It is believed that the higher the copy number is, the more efficient the plasmid is at replicating itself.
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.
What is the function of the plasmid?
Plasmids are extra circular genetic material that can be passed from bacteria to bacteria, which basically is their function; in bacterial conjugation. But, in biotechnology it is often used in recombination work. Some other organisms gene is inserted into the bacterial plasmid and then the bacteria multiply and transcribe this inserted gene into many useful products.
How can bacteria be transformed with recombinant plasmid?
Bacteria can be transformed with recombinant plasmid by introducing the plasmid into the bacterial cell through a process called transformation. This allows the bacteria to take up the recombinant DNA from the plasmid and express the desired gene or trait encoded in the DNA.
