Recombinant DNA technology involves the creation of new genetic combinations by combining DNA from different sources. It is also known as rDNA technology.
The process includes various steps, starting from isolating the desired gene. It proceeds further by cutting DNA with restriction enzymes and inserting the gene into a vector.
The end of the process is marked by the introduction of the vector into host cells and screening for successful gene integration.
In this blog, we will understand the steps involved in Recombinant DNA technology and its applications.
About Recombinant DNA Technology
Recombinant DNA technology is also known as rDNA technology or genetic engineering. The technique has a crucial role in biology. It is about manipulating the DNA fragments of an organism and modifying them.
The technique involves the altering of one organism’s DNA sequence to attain specific outcomes. It also helps in the creation of new genetic combinations.
The steps required in the technique involve the use of enzymes and various lab techniques. The enzymes and techniques help manipulate the DNA segments and join them.
Recombinant DNA technology has been a breakthrough in biology and biotechnology applications. It supports scientific studies and various types of research.
Technology facilitates genetic studies and enables new inventions in pharmaceuticals, and agricultural applications.
Steps Involved In Recombinant DNA Technology
In Recombinant DNA technology, the recombinant molecules are created to introduce them into host cells. The steps involved in this process are as follows:
- Genetic Material Isolation – The DNA segments of interest are treated with several enzymes and some lab techniques. They help in the isolation of the DNA by cutting the fragments using restriction enzymes.
Restriction enzymes serve as the molecular scissors that cut the DNA at specific regions. The DNA fragments obtained after cutting will be combined with the DNA of another species.
Isolation of DNA is done using various techniques such as cell lysis and enzymatic treatments, which release the DNA from the cells. The recombination method allows the creation of new genes with new functions.
- Vector Preparation – In this step, an appropriate vector, such as plasmid, bacteria, or virus is chosen to carry the gene. The vector serves as a medium to pass the genes to the body of another organism.
- Introduction Of Vector into Host Cells – Again, restriction enzymes cut the vector, to create compatible ends for inserting the genes into the vector.
The DNA fragments attach to vector DNA with the help of DNA ligase, which is an enzyme. The function of restriction enzyme is to separate the DNA fragments, while DNA ligase joins the DNAs of two organisms.
- Choosing The Transformed Cells – Host cells may not accept the transfer of all recombinant DNA. It is essential to identify the cells that have received the recombinant DNA successfully.
To identify cell transformation, selectable markers such as antibiotic-resistance genes are used. These markers are present in the vector.
- Gene Cloning and Expression of Recombinant DNA – The cells receiving the recombinant DNA are cultured to multiply and produce copies of themselves. All the clones will contain recombinant DNA.
To confirm the presence of specific genes and expressions, copies are screened. The techniques used for cloning procedures include PCR, Southern blotting, and sequencing.
- Extraction of Recombinant Protein – Recombinant proteins are the product of recombinant DNA technology. To produce a recombinant protein, the host cells are induced by gene expression.
Further, the protein is extracted, and its purification is done using various techniques such as chromatography.
- Analysis of Recombinant DNA and Protein – DNA analysis is the final step that involves analyzing the recombinant DNA and protein. Analysis is done to ensure the resulting DNA is precise and functional.
This can involve various biochemical and molecular biology techniques to verify the structure and activity of the recombinant product.
The Link Between DNA Drawing and Recombinant DNA Technology
DNA drawing is a technique that involves a visual representation of DNA sequences and structures. The method helps in understanding and designing genetic alterations in recombinant DNA technology.
DNA drawing enables the process of introducing new genetic combinations into host cells. This allows the use of the technology in various applications requiring research and biotechnology.
Use Of Microsatellite DNA In Recombinant DNA Technology
Microsatellite DNA is a of short, repetitive segments of DNA. These are scattered throughout the genome. Microsatellite DNA is used in recombinant DNA technology as a genetic marker for the identification and mapping of genes.
Microsatellite DNA also aids in studying genetic diversity and tracing inheritance patterns. Its use increases the precision and success of genetic engineering and molecular biology research.
Recombinant DNA Technology and Genetic Testing
Recombinant DNA or rDNA technology plays a significant role in genetic testing. It enables the precise manipulation and accurate analysis of genetic material.
The process of Genetic testing involves DNA analysis for the identification of changes or mutations. Such changes in the genetic material may potentiate the risk of genetic disease or specific traits.
Recombinant DNA technology creates recombinant DNA molecules. Researchers can perform amplification of DNA segments to determine genetic mutations or variants. Gene mutations can be associated with hereditary disorders.
The technology also aids in developing DNA probes and markers for precise detection. This helps construct genetic libraries for accurate gene identification, enabling the production of recombinant proteins for diagnostic evaluations.
The use of rDNA technology supports various applications, enhancing the accuracy and scope of genetic testing. The technique facilitates effective diagnosis, and plans treatment strategies. It also allows a deeper understanding of genetic disorders and associated risks.
Conclusion
The steps involved in Recombinant DNA technology or rDNA technology perform precise functions with multiple sub-steps.
Each of the sub-steps has specific protocols that may vary according to the organisms and preferred vectors.
The set of protocols will also differ based on the specific goals of the recombinant DNA technology application.
The use of rDNA technology improves sequencing methods. It obtains detailed genetic information that can be further used for diagnostic purposes, personalized medications, and improved success outcomes.
