A vaccine is a type of microorganism made from a suspension of weakened, semi-dead or dead micro-organisms that is used to prevent attacks by similar micro-organisms. Edward Jenner discovered smallpox and Louis Pasteur discovered the rabies vaccine. It is of four types. Live vaccine, Killed vaccine, Toxoid vaccine and breed based vaccine.
Importance, function of interferon
1. It inhibits the growth of virus.
2. It regulates the immune system.
3. | Prevents the increase in the number of B and T lymphocytes. Cures viral diseases.
4. Prevents the growth of cancer cells by increasing the capacity and number of Natural Killer Cells.
5. Destroys cancer cells by increasing the host’s immunity.
6. It inhibits the production of antibodies. Stimulates innate antioxidant capacity and reproduction.
7. Interferon is used to treat complicated hepatitis-B, herpes, papilloma etc.
8. It is effective against the rabies virus.
Steps in interferon production
1. Target gene selection and isolation: DNA is collected from human fibroblast cells. The gene carrying the interferon code (interferon-beta) is isolated from the DNA.
2. Carrier Selection: Carriers are selected to carry the target gene. Plasmids act as DNA carriers.
3. Local Excision of Plasmid DNA: Specific regions are excised from the carrier plasmid DNA using restriction enzymes.
4. Preparation of recombinant DNA: The interferon gene is ligated to the carrier plasmid DNA by ligase enzymes. As a result, recombinant DNA is produced.
5. Introduction of Recombinant DNA into Host: Recombinant DNA is introduced into cells of host E. coli bacteria. The process of introducing recombinant DNA into bacterial cells is called transformation.
6. Amplification of Recombinant DNA: After the recombinant DNA is introduced into the host cells, the host bacteria are cultivated in culture medium. Within a short period of time, the bacteria multiply in the culture medium to produce thousands of copies. Recombinant DNA is also produced. During this time the bacteria secrete interferon through the culture.
7. Isolation of Interferon: Interferon is isolated from the culture medium. It is then purified by chemical means.
Each yeast cell produces one million (10 lakh) molecules of interferon. 1×105 molecules of interferon are produced inside E. coli.
What are Interferons?
Interferon is a high molecular weight protein. Small groups of proteins with a molecular weight of 20,000-30,000 daltons are called interferons. It is a protective protein. Interferon is produced by E. coli and yeast. The commercial name of interferon is Betaferon. It is of three types. α-interferon β-interferon and γ-interferon.
In 1957, British scientists Alick Isaacs and Jean Lindermann discovered interferon. In 1980, American scientists Gilbert and Weissmann (Gilbert & Weissmann) were able to produce human interferon.
Steps in the insulin production process
1. Synthesis of Polynucleotide Chains: Insulin molecules are composed of two polynucleotide chains. A chain and B chain. A chain consists of 21 and B chain of 30 amino acids. A chain of 63 nucleotides and B chain of 90 nucleotides are made chemically in the laboratory. Then the two chains are purified.
2. Production of recombinant DNA: Carrier bacteria are selected. Plasmid DNA is isolated from bacterial cells. Restriction enzymes are used to remove certain parts from A chain and B chain. Again, corresponding segments are cut from plasmid DNA using the same enzyme. However, the portion of the plasmid DNA that produces the gene for the β-galactosidase enzyme is cut out. The A chain and B chain are then joined to separate plasmid DNA with the help of ligase enzyme. Recombinant DNA is formed as a result of ligation of A chain and B chain with plasmid DNA. Thus two types of recombinant DNA are produced. A strand is recombinant DNA and B strand is recombinant DNA.
3. Introduction of Recombinant DNA into Host: Recombinant DNA is introduced into host bacterial cells during the transformation process. This is done by heat-shock method or electric pulse method. The host (E. coli) is first placed in CaCl2 solution and then on ice for 14-16 hours. It binds Ca to the host cell wall. A mixture of host and recombinant DNA in a container was placed on ice for 30 min, at 420°C for 90 s, and again on ice for 2 min. In this the host (E. coli) absorbs the recombinant DNA.
4. Gene Cloning: Host bacteria (E. coli) with recombinant DNA are grown in culture medium in fermentation tanks. As the number of E. coli bacteria increases, recombinant DNA copies are also produced. This results in β-galactosidase A chain and β-galactosidase B chain. Also some methionine is produced.
5. Separation of the two chains: A chain and B chain of the polynucleotide are separated from the culture medium. The resulting methionines with the A chain and B chain are then removed using cyanogen bromide. The two chains are then chemically purified.
6. Preparation of Insulin Molecule: In sulfonating process, A chain and B chain of nucleotides are combined in presence of sodium disulfonating and sodium sulfite. The two chains are then recombined by a disulfide bond.
7. Purification of insulin: The insulin produced contains the bacteria’s own protein. Therefore the extracted insulin is chemically purified.
8. Insulin Market Types: Pure insulin is filled in suitable ampoules. Then it is marketed.
Insulin discovery and research
In 1916, Sir Edward Sharpe Shaffer discovered insulin secreted by the human pancreas. In 1921, Canadian physiologists Banting and Best described the antidiabetic role of insulin. In 1923, Eli Lilly collected and marketed insulin from the pancreas of pigs and cows. In 1954, Frederick Sanger discovered the amino acid sequence and chemical structure of human insulin. In 1977, Itakura and his colleagues first synthesized two chains of human insulin. In 1980, Novo Industry of Denmark converted pig insulin into 99% pure human insulin. In 1981, scientists at Hope National Medical Center in California produced pure human insulin. In 1982, Eli Lilly commercially produced human insulin called Humulin. Currently, companies such as Genintech, Novo Industries, Biogen, Eli Lilly etc. are commercially marketing pure human insulin.
What is insulin?
Insulin is a type of hormone. Insulin is the hormone released from the beta (β) cells of the pancreas that controls diabetes by accelerating the uptake of blood glucose. Insulin is a type of small protein. Pro-insulin consists of four parts. Anterior part and A, B and C parts. Insulin is made by removing the C portion of pro-insulin and joining the A and B portions by a disulfide bond. Human insulin contains 17 types of 51 amino acids. Its chemical symbol is C254H377N65O75S6 and molecular mass is 5734. The DNA molecule at the top of the short arm of human chromosome 11 contains the insulin producing gene. It consists of 153 nitrogenous bases. E. coli bacteria contain 1×105 (10 lakh) insulin molecules.
What is anoxia?
Anoxia happens when body or brain completely loses its oxygen supply. Anoxia is usually a result of hypoxia. This means that a part of body doesn’t have enough oxygen. When body is harmed by a lack of oxygen, it’s called a hypoxic-anoxic injury.
Symptoms of anoxia
- Mood and personality changes
- Memory loss
- Slurred speech or forgotten words
- Changes in judgment
- Trouble walking or moving arms or legs normally
- Weakness
- Feeling dizzy or disoriented
- Unusual headaches
- Trouble concentrating
Limitations of genome sequencing in Bangladesh
1. Lack of laboratories: Genome sequencing experiments are conducted in various universities, medical colleges, biotechnology institutes, government and private clinics in Bangladesh. But its number is much less than the requirement. Therefore, there is a need to increase the number of research institutes in public, private and private sectors.
2. Scarcity of research equipment: Most of the equipment and materials for genome sequencing experiments are imported from abroad. Therefore, the necessary equipment and materials are not sufficient. Many appliances have been rendered useless. These machines and materials should be produced through indigenous technology. 3. Limited Skilled Researchers: The number of skilled researchers in genetics and genome sequencing research in Bangladesh is much less than required. Many private or private medical institutions do not have skilled researchers.
4. Lack of practical training: Regular training of doctors and researchers needs to be arranged. Regular training is the best tool for efficiency. Therefore practical training should be arranged in public and private ways.
5. Lack of skilled assistants: Most of the assistants employed in research work are unskilled.
6. Financial constraints: Genome sequencing research requires expensive laboratories, skilled researchers and skilled assistants. This requires a lot of money. The public and private funds provided are not sufficient.
Prospects of genome sequencing in Bangladesh
1. Applications in paternity determination: Genome sequencing is being successfully applied to identify the father or mother of a fetus. A child inherits half of its chromosomes from its parents. So paternity is being determined by identifying the child’s DNA. This is one of the breakthroughs in genome sequencing.
2. Application in Criminal Identification: Criminals involved in murder or rape are identified through the application of genome sequencing. Blood drops, hair, teeth, nails, semen or semen, body parts etc. are collected from the crime scene. If the amount of sample obtained is too low, the amount is amplified in the PCR process. The genome sequencing of these samples is compared with the genome sequencing of the suspect. This is how criminals are identified.
3. Gene therapy and RNAi: Cure genetic defects using gene therapy. By removing the defective gene from the cell and inserting the healthy gene, the patient is cured of the disease. Viruses, RNAi, antisense or zinc finger proteins are used as carriers of these genes. As a result, the treatment of hemophilia, color blindness, night blindness, cancer etc. has become easier.
4. Stem cells: Cells that can divide for life are called stem cells. B-lymphocytes and T-lymphocytes are stem cells. These cells are used to replace lost organs, make organ cells and test new drugs.
5. Micro RNA: Micro RNA can be used in the diagnosis and treatment of diseases such as cancer, viral infections, metabolic disorders, inflammation etc. It inhibits the action of disease-causing genes.
6. Genome Scanning: Genome scanning technology allows the complete genome of any organism to be known at low cost and very quickly. In this process, it will be possible to uncover information about 98% of inactive genes in human chromosomes.
7. Nanotechnology: The provision of human health services and food security will be possible through the production of organic matter through nanotechnology.
8. Gene Cloning: Many materials used for human welfare are created by applying gene cloning technology. In the future it will be possible to produce essential items using this technology.
9. GM micro-organisms: In the developed countries of the world, using GM micro-organisms is trying to prevent the level of environmental pollution. GM microorganisms will be used to keep the natural environment clean.
10. Research Institutions: Genome sequencing experiments are conducted in various universities, medical colleges, biotechnology institutes, government and private clinics in Bangladesh. The number of research institutes is increasing in public, private and private endeavors. As a result, the potential for genome sequencing is increasing.
11. Research Materials: Most of the equipment and materials for genome sequencing experiments are imported from abroad. So the cost becomes high. At present several machines and materials are being produced in the country.
12. Skilled Researchers: Bangladesh has skilled researchers for research on genome sequencing and genetics. Researchers are being recruited all the time.
13. Training Arrangements: Regular training is arranged for people engaged in genome sequencing and genetics research work in the country and abroad. As a result, the possibility of genome sequencing in Bangladesh is progressing.