Biotechnology combines biology and technology for the benefit of humanity and long-term development. Bioinformatics, bioprocess engineering, and genetic engineering are part of modern biotechnology.
We can change an organism’s genetic composition through genetic engineering techniques. We insert Genes of choice, such as pest-resistant or antibiotic-resistant genes, into a host organism using a cloning vector. It integrates the desired gene into the host genome, transforming the host’s phenotype.
With bioprocess engineering, you can make various products such as enzymes and antibodies on a much larger scale. We grow the desired microorganisms in controlled, sterile conditions.
Following are the principles of biotechnology:
Modern biotechnology is the outcome of the following two basic principles and processes of biotechnology.
It includes techniques for modifying genetic material, such as RNA or DNA, to introduce it into host organisms. It involves phenotypic changes in the host.
Maintaining microbial contamination-free environments in chemical engineering processes maximises cell growth and produces biotechnological products such as enzyme production, vaccine production, and antibiotic production.
Concepts for genetic engineering conceptual development
Sexual reproduction is better than asexual reproduction. It is because sexual reproduction enables the creation of new genetic setups and enables variation, whereas a sexual reproduction aims to preserve existing genetic information. This concept led to gene transfer, recombinant DNA, and gene cloning, which have major significance in genetic engineering. These methods have enabled the isolation of the desired gene without introducing an undesirable one into the target organism.
We can link an antibiotic resistance gene to a Salmonella Typhimurium plasmid using a synthetic recombinant DNA molecule. Herbert Boyer and Stanley Cohen isolated the antibiotic resistance gene from plasmid DNA in 1972. The discovery of molecular scissors enabled this type of DNA cutting. After cutting the DNA, it connected to plasmid DNA, which served as vectors to transfer the DNA attached to it. An enzyme known as DNA ligase has made this linking possible, enabling the development of recombinant DNA in vitro.
Tools of recombinant DNA technology
Scientists discovered two enzymes in 1963 to help control bacteriophage growth in Escherichia coli, the first of which added methyl groups to DNA and used them to cut DNA.
After recognising the sequenced base pairs, the first restriction endonuclease, Hind II, depended on the precise sequence of DNA nucleotides to cut DNA molecules at a specific point. The recognition sequence refers to this particular sequence of base sequences. We now have around 900 restriction enzymes from over 30 bacterial strains with Hind II, and these enzymes have different recognition sequences.
Restriction enzymes are part of a larger group of enzymes known as nucleases, further divided into endonucleases and exonucleases. Endonucleases cut DNA, whereas exonucleases assist in removing nucleotides from DNA.
Because each restriction endonuclease inspects the length of the DNA sequence, the enzyme EcoRI is vital in the creation. When the specific recognition sequence is present, it binds with DNA and cuts each strand of the double helix at a specific sugar-phosphate backbone.
DNA fragment separation and isolation
Restriction endonucleases separate DNA fragments using gel electrophoresis. We move negatively charged DNA fragments towards the anode via a matrix or other medium. The most prevalent matrix is agarose, a naturally occurring polymer from seaweeds. Agarose gel’s sieving effect separates DNA fragments based on their size.
Features required facilitating cloning into a vector
Origin of replication
This sequence starts replication and all DNA links to it within the host cell. Because the origin of replication helps control the copy number of linked DNA, if someone wants to recover multiple copies of target DNA, we must clone it in a vector with a high copy number origin.
Selectable markers are necessary to eliminate and identify non-transformants and enable the growth of transformants selectively. The process of introducing a new DNA piece into the host bacterium is transformation.
The restriction enzyme uses recognition sites to link alien DNA. When the vector contains more than one recognition site, it generates multiple fragments, complicating gene cloning.
Animal and plant cloning vectors
Viruses and bacteria transmit genes in plants and animals. For example, the pathogen Agrobacterium tumefaciens can use T-DNA to transform normal plant cells into tumour cells that produce the pathogen’s required chemical. In animals, retroviruses can transform normal cells into cancerous cells.
DNA cannot cross the cell membrane because it is a hydrophilic molecule. Bacteria must first be “competent” to take up the plasmid by treating them with a specific divalent cation concentration like calcium and incubating cells on ice with recombinant DNA forces recombinant DNA into the cells.
It is not the only method of introducing alien DNA into host cells; micro-injection injects alien DNA into animal cells’ nuclei.
Recombinant DNA Technology process
Isolation of DNA
Because membranes consist of DNA, it is necessary to break the cells open to release DNA and RNA, proteins, lipids, and polysaccharides. We carry out isolation in the presence of several enzyme-producing agents, such as cellulose (plant cells), chytinase (bacteria), and lysozyme (fungus).
DNA cutting at specific locations
We can perform restriction enzyme digestion by incubating a purified DNA molecule with a restriction enzyme under ideal conditions. Agarose gel electrophoresis monitors the progress of enzyme digestion. Because DNA has a negative charge, it attracts positive electrodes, and we repeat the process with the vector DNA.
PCR Amplification of Gene of Interest
PCR uses two sets of primers to make numerous copies of a gene (or DNA) in vitro. Nucleotide-extending enzymes extend primers and use genomic DNA as a template. Repeating DNA replication results in a billion times amplification of a particular DNA segment, and Thermostable DNA polymerase repeats the amplification.
Recombinant DNA insertion in host cells/organisms
We use several methods to introduce litigated DNA into recipient cells capable of receiving and taking up DNA from their surroundings.
Getting the foreign gene product
The alien DNA multiplies when inserted into the cloning vector. It is essential to express recombinant DNA in most recombinant technologies to produce desired proteins. We can express the foreign gene in host cells under the right conditions, but it requires a lot of technical details.
After the biosynthetic stage, we separate and purify the product to form the finished product before marketing. There are various preservatives used in these products, and each has different processes and quality control testing.
It is the branch of biology that uses both technology and living things. The field of biotechnology deals with the creation, modification, and distribution of products that benefit human health and well-being. It’s one of the oldest known industrial technologies. Fermentation in the manufacturing of alcoholic beverages is a great example of biological technology. We apply this biological technique in various fields, including genetic engineering, agriculture, and medicine.
Bioinformatics, a branch of biotechnology, is useful for researchers and developers. We use biochemical engineering to extract and create living entities due to this research.
Medicine, genetics, Agriculture, and numerous commercial products such as alcohol, wine, and chemicals use biotechnology.