Nimona Genjosa
Aug 5
List & explain methods of introduction of Recombinant DNA molecules mechanisms into the host cell?
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Aug 5
1. Transformation: In this method, the host cell is made competent to take up the recombinant DNA molecules by treating it with calcium chloride or electroporation. The recombinant DNA is then added to the competent cells, which can take up the DNA and incorporate it into their genome.
2. Transfection: Transfection involves the introduction of recombinant DNA molecules into eukaryotic cells using methods such as calcium phosphate precipitation, electroporation, or lipofection. The DNA can then be integrated into the host cell's genome or remain as an episome.
3. Electroporation: In electroporation, a brief electric pulse is applied to the host cell, creating temporary pores in the cell membrane through which the recombinant DNA molecules can enter. This method is commonly used for introducing DNA into bacterial cells.
4. Microinjection: Microinjection involves directly injecting recombinant DNA molecules into the nucleus of a eukaryotic cell using a fine needle. This method is labor-intensive and requires specialized equipment but can be used to introduce DNA into specific cells.
5. Viral vectors: Viral vectors can be used to introduce recombinant DNA molecules into host cells. The DNA is inserted into the viral genome, and the virus is then used to infect the host cell, delivering the DNA into the cell's genome.
6. Liposome-mediated transfection: Liposomes are lipid vesicles that can encapsulate and deliver DNA molecules into cells. The DNA-liposome complex is taken up by the host cell through endocytosis, and the DNA is released into the cytoplasm.
7. Biolistic particle delivery: In this method, DNA-coated gold or tungsten particles are shot into the host cell using a gene gun. The DNA is released into the cell's cytoplasm or nucleus upon impact, allowing for the integration of the recombinant DNA into the host cell's genome.
2. Transfection: Transfection involves the introduction of recombinant DNA molecules into eukaryotic cells using methods such as calcium phosphate precipitation, electroporation, or lipofection. The DNA can then be integrated into the host cell's genome or remain as an episome.
3. Electroporation: In electroporation, a brief electric pulse is applied to the host cell, creating temporary pores in the cell membrane through which the recombinant DNA molecules can enter. This method is commonly used for introducing DNA into bacterial cells.
4. Microinjection: Microinjection involves directly injecting recombinant DNA molecules into the nucleus of a eukaryotic cell using a fine needle. This method is labor-intensive and requires specialized equipment but can be used to introduce DNA into specific cells.
5. Viral vectors: Viral vectors can be used to introduce recombinant DNA molecules into host cells. The DNA is inserted into the viral genome, and the virus is then used to infect the host cell, delivering the DNA into the cell's genome.
6. Liposome-mediated transfection: Liposomes are lipid vesicles that can encapsulate and deliver DNA molecules into cells. The DNA-liposome complex is taken up by the host cell through endocytosis, and the DNA is released into the cytoplasm.
7. Biolistic particle delivery: In this method, DNA-coated gold or tungsten particles are shot into the host cell using a gene gun. The DNA is released into the cell's cytoplasm or nucleus upon impact, allowing for the integration of the recombinant DNA into the host cell's genome.
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Google Gemini AI
Aug 5
Methods of Introduction of Recombinant DNA Molecules into Host Cells
1. Transformation:
- Bacterial cells are treated with calcium chloride (CaCl2) to make them permeable to DNA.
- Recombinant DNA is mixed with the competent bacterial cells and left to incubate.
- DNA enters the cells through the permeable cell membrane.
- Efficiency is relatively low, but it is a simple and cheap method.
2. Electroporation:
- Cells are exposed to a brief electrical pulse in the presence of recombinant DNA.
- The electrical pulse creates transient pores in the cell membrane, allowing DNA to enter.
- Efficiency is higher than transformation but requires specialized equipment.
3. Microinjection:
- A glass capillary needle is used to inject DNA directly into the nucleus of individual cells.
- High efficiency, but it is a labor-intensive method and can damage cells.
4. Biolistics (Particle Bombardment):
- Tiny DNA-coated gold or tungsten particles are shot into cells using a particle gun.
- DNA enters cells through the holes created by the particles.
- Suitable for plant cells and some other difficult-to-transform cells.
5. Viral Vectors:
- Viruses are used as vectors to deliver recombinant DNA into host cells.
- The viral genome is modified to carry the desired gene, and the virus is then used to infect host cells.
- High efficiency, but viruses can be associated with safety concerns.
6. Transfection:
- Calcium phosphate, DEAE-dextran, or lipofectamine are used to create lipid-DNA complexes that facilitate the entry of DNA into eukaryotic cells.
- Efficiency is variable and depends on cell type and transfection reagents.
7. Retroviral Vectors:
- Retroviruses are engineered to carry the desired gene.
- When the retrovirus infects a host cell, its RNA genome is reverse transcribed into DNA and integrated into the host cell's genome.
- High efficiency, but retroviruses have safety concerns and can only infect dividing cells.
1. Transformation:
- Bacterial cells are treated with calcium chloride (CaCl2) to make them permeable to DNA.
- Recombinant DNA is mixed with the competent bacterial cells and left to incubate.
- DNA enters the cells through the permeable cell membrane.
- Efficiency is relatively low, but it is a simple and cheap method.
2. Electroporation:
- Cells are exposed to a brief electrical pulse in the presence of recombinant DNA.
- The electrical pulse creates transient pores in the cell membrane, allowing DNA to enter.
- Efficiency is higher than transformation but requires specialized equipment.
3. Microinjection:
- A glass capillary needle is used to inject DNA directly into the nucleus of individual cells.
- High efficiency, but it is a labor-intensive method and can damage cells.
4. Biolistics (Particle Bombardment):
- Tiny DNA-coated gold or tungsten particles are shot into cells using a particle gun.
- DNA enters cells through the holes created by the particles.
- Suitable for plant cells and some other difficult-to-transform cells.
5. Viral Vectors:
- Viruses are used as vectors to deliver recombinant DNA into host cells.
- The viral genome is modified to carry the desired gene, and the virus is then used to infect host cells.
- High efficiency, but viruses can be associated with safety concerns.
6. Transfection:
- Calcium phosphate, DEAE-dextran, or lipofectamine are used to create lipid-DNA complexes that facilitate the entry of DNA into eukaryotic cells.
- Efficiency is variable and depends on cell type and transfection reagents.
7. Retroviral Vectors:
- Retroviruses are engineered to carry the desired gene.
- When the retrovirus infects a host cell, its RNA genome is reverse transcribed into DNA and integrated into the host cell's genome.
- High efficiency, but retroviruses have safety concerns and can only infect dividing cells.
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