Steps of Creating a transgenic plant in lab

Creating a transgenic plant involves several steps in the laboratory using modern biotechnology techniques. Below is an overview of the process to create a transgenic plant:-

1. 1. Identify the Desired Gene: The first step is to identify and isolate the gene with the desired trait that you want to introduce into the target plant. This gene could come from another plant species, animals, bacteria, or any other source.

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3. 2. Prepare the Gene of Interest: Once the desired gene is identified, it needs to be isolated and amplified through techniques like polymerase chain reaction (PCR) to create multiple copies of the gene.

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5. 3. Construct the Plant Expression Vector: The isolated gene is then inserted into a plant expression vector. A plant expression vector is a small circular piece of DNA that acts as a carrier to transfer the gene into the plant cells. The vector contains regulatory sequences that control the expression of the gene, ensuring it functions correctly in the plant.

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7. 4. Introduce the Vector into Plant Cells: The constructed plant expression vector is now introduced into plant cells. There are various methods to achieve this, including:

   a. Agrobacterium-mediated transformation: Using a soil bacterium called Agrobacterium tumefaciens, the vector is transferred into the plant cells naturally infected by the bacterium.

   b. Biolistic (or gene gun) method: Tiny gold or tungsten particles coated with the plant expression vector are shot into plant cells using a gene gun.

   c. Electroporation: Applying a brief electric shock to the plant cells to create temporary pores through which the vector can enter.

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9. 5. Regenerate Transgenic Plants: Once the plant cells have taken up the vector and integrated the foreign gene into their genome, they are grown on a special medium to develop into transgenic plantlets. The plantlets are then transferred to the soil to grow into mature transgenic plants.

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11. 6. Screening and Selection: Not all plant cells will successfully incorporate the foreign gene. Therefore, the transformed plant cells need to be identified and selected from the non-transformed ones. Selective markers (e.g., antibiotic or herbicide resistance genes) are often included in the plant expression vector to aid in this process. Only the transformed cells will survive in the presence of the marker.

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13. 7. Confirmation of Transgene Integration: The presence and stability of the transgene in the regenerated plants need to be confirmed. Techniques like PCR and Southern blotting are used to verify the presence and copy number of the introduced gene.

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15. 8. Testing and Characterization: The transgenic plants with confirmed gene integration are subjected to extensive testing and characterization to assess the expression of the desired trait and to ensure there are no unintended effects.

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17. 9. Field Trials and Regulatory Approval: Before commercial use, transgenic plants usually undergo field trials to assess their performance and potential environmental impact. Regulatory authorities review the safety and efficacy data to grant approval for commercial cultivation.

It is crucial to mention that creating transgenic plants requires specialized laboratory facilities, skilled researchers, and adherence to strict biosafety protocols and regulatory guidelines. Moreover, potential environmental and ethical concerns should be addressed during the development and commercialization of transgenic plants

what is transgenic plant,

 A transgenic plant is a genetically modified organism (GMO) created by introducing genes from one species into the genome of a plant. The process of creating a transgenic plant involves the use of recombinant DNA technology, where specific genes from one organism are isolated and inserted into the genetic material of the target plant.

The introduction of these new genes can serve various purposes, including:

1. 1. Improved Crop Traits: Transgenic plants are often designed to express desirable traits, such as increased resistance to pests, diseases, or environmental stresses like drought or extreme temperatures. For example, a gene from a bacterium might be inserted into a crop plant to make it resistant to certain insects.

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3. 2. Enhanced Nutritional Content: Genes can be added to plants to increase their nutritional value. For instance, scientists have developed transgenic plants with higher levels of essential vitamins or nutrients.

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5. 3. Production of Pharmaceuticals: Some plants are engineered to produce pharmaceutical compounds, vaccines, or other medical products. These plants are often called "biopharmaceuticals" or "pharmaceutical crops."

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7. 4. Environmental Benefits: Transgenic plants can also be designed for environmental purposes, such as phytoremediation. This process involves using plants to remove pollutants from the soil or water.

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9. 5. Research and Development: Transgenic plants are used in research to study gene functions and understand the underlying mechanisms of various biological processes.

It is essential to note that the development and use of transgenic plants have raised ethical, environmental, and regulatory concerns. Critics argue about potential risks to the environment and the need for rigorous safety assessments before releasing transgenic plants into the wild or commercial use.

The creation and use of transgenic plants are subject to strict regulations in many countries to ensure the safety of the environment, human health, and biodiversity. Different regions have different policies and guidelines for the development and commercialization of GMOs, including transgenic plants.

what is translation in biology,

In biology, translation is a crucial process that occurs within cells, where genetic information encoded in messenger RNA (mRNA) is used to synthesize proteins. This process is essential for the proper functioning of living organisms because proteins play various critical roles in cell structure, function, and regulation.

Here's a simplified explanation of translation:-

1. 1. DNA Transcription: The process begins with the transcription of a gene in the DNA into a complementary mRNA molecule. This mRNA molecule carries the genetic information from the DNA to the ribosomes, the cellular machinery responsible for protein synthesis.

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3. 2. mRNA Processing: Before the mRNA leaves the cell nucleus and enters the cytoplasm where ribosomes are located, it undergoes some modifications like the removal of introns (non-coding regions) and the addition of a 5' cap and a 3' poly-A tail.

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5. 3. Initiation: The mRNA binds to the small ribosomal subunit in the cytoplasm. This complex then attaches to the start codon (AUG) on the mRNA, which signals the beginning of the protein-coding sequence.

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7. 4. Elongation: The ribosome moves along the mRNA, reading the codons (groups of three nucleotides) in the mRNA sequence. Each codon codes for a specific amino acid. Transfer RNA (tRNA) molecules bring the corresponding amino acids to the ribosome, where they are linked together in a growing polypeptide chain.

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9. 5. Termination: The process continues until the ribosome reaches a stop codon (UAA, UAG, or UGA) in the mRNA sequence. These codons do not code for any amino acids but signal the end of translation. At this point, the newly synthesized polypeptide is released from the ribosome.

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11. 6. Protein Folding: After translation, the polypeptide chain undergoes folding to acquire its three-dimensional shape, which is critical for its proper function.

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13. 7. Post-Translational Modifications: In some cases, the protein may undergo additional modifications after translation, such as the addition of chemical groups or cleavage of certain sections to become fully functional.

The entire process of translation is tightly regulated and precise, ensuring that the correct proteins are produced in the right quantities to carry out various cellular functions. Any errors or mutations during translation can lead to significant biological consequences and may cause various diseases or disorders.

what is transcription in biology

In biology, transcription is a fundamental process that involves the synthesis of RNA molecules from a DNA template. It is a crucial step in gene expression and is responsible for converting the genetic information stored in DNA into functional RNA molecules that can perform various cellular functions.

The process of transcription is carried out by a large enzyme called RNA polymerase. Here's a brief overview of the transcription process:

1. Initiation: Transcription begins with the binding of RNA polymerase to a specific region of the DNA called the promoter. The promoter region is located upstream (towards the 5' end) of the gene being transcribed. Once the RNA polymerase is bound to the promoter, it unwinds a small portion of the DNA, exposing the template strand.

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3. Elongation: As the RNA polymerase moves along the DNA template strand, it synthesizes an RNA molecule that is complementary to the DNA sequence. The RNA polymerase reads the DNA template in the 3' to 5' direction and synthesizes the RNA molecule in the 5' to 3' direction. The RNA molecule is assembled using ribonucleotides (A, U, G, and C), and the base-pairing rules are the same as in DNA: A pairs with U (in RNA) and G pairs with C.

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5. Termination: Transcription continues until the RNA polymerase encounters a termination sequence on the DNA template. This sequence signals the RNA polymerase to stop synthesizing the RNA molecule and release it. In some cases, termination is dependent on specific proteins that help release the RNA polymerase from the DNA.

Once transcription is complete, the newly synthesized RNA molecule, known as messenger RNA (mRNA), undergoes further processing in eukaryotic cells before it can be used to produce proteins. This processing involves the addition of a 5' cap and a poly-A tail to the mRNA, as well as the removal of non-coding regions (introns) through a process called splicing. In prokaryotic cells, the mRNA is often ready for translation into proteins without extensive processing.

In summary, transcription is the process by which RNA molecules are synthesized from a DNA template, and it plays a central role in the flow of genetic information from DNA to functional proteins in the cell.

difference between Innate and adaptive immunity

 

Analytical/Preparative Centrifugation

 Analytical/Preparative Centrifugation:-

The 2 most common types of centrifugation are analytical and preparative; the distinction is between the 2 is based on the purpose of centrifugation. Analytical centrifugation involves measuring the physical properties of the sedimenting particles, such as sedimentation coefficient or molecular weight. Optimal methods are used in analytical ultracentrifugation. Molecules are observed by optical system during centrifugation, to allow observation of macromolecules in solution as they move in the gravitational field. The samples are centrifuged in cells with windows that lie parallel to the plane of rotation of the rotor head. As the rotor turns, the images of the cell (proteins) are projected by an optical system onto film or a computer. The concentration of the solution at various points in the cell is determined by absorption of a light of the appropriate wavelength. This can be accomplished either by measuring the degree of blackening of a photographic film or by the deflection of the recorder of the scanning system and fed into a computer. The other type of centrifugation is called preparative and the objective is to isolate specific particles that can be reused. There are many type of preparative centrifugation such as rate zonal, differential, and isopycnic centrifugation.