Molecular biology began more than 40 years ago with Watson and Crick’s discovery of the double helix structure of deoxyribonucleic acid DNA and that continued with Cohen and Borer’s. This discovery of the techniques that called as biotechnology nowadays has had an enormous impact on the nature of research and development. This technique has potential to generate great benefits in improved of human life by through of many applications which play role in improvements of medicine, agriculture, food production, environment and others.However, in this article, only lath and agriculture sectors which developed from molecular biology techniques were discussed.
Recombinant DNA technology is the one that is widely used in biotechnology fields which is resulting recombinant of proteins and other products that result from the use of radar that is found in essentially on every health and agriculture sector. The most common application of recombinant DNA technologies that was used in health sector for example are human insulin and hepatitis B vaccine.Recombinant DNA human insulin developed by molecular biology technique gives huge contribution to health sector in order to produce biosynthesize insulin rotten which helps control level of sugar in diabetes patients, who are disable or do not correctly produce their insulin from their body. Insulin is the internal secretion of the pancreas formed by groups of cells called the islets of Lanterns. It is the hormone needed to enable glucose to enter the cells and provide energy and also important in keeping blood glucose levels within the acceptable limits.The biosynthesize insulin which produced through recombinant DNA technology is injected into the body by people who suffered from diabetes with type 1 diabetes in whom the cells that produce insulin have been destroyed.
This is the most common form of diabetes in children and young adults, and they depend on insulin for survival. Insulin may also be used by people with type 2 diabetes, where the body needs more insulin than it can produce.Early preparations of insulin were purified quite crudely from pancreas tissue extracted from animals either pigs or cattle.
However, insulin is mostly made pessimistically nowadays by recombinant DNA technology or ‘genetic engineering’. While, for the recombinant DNA hepatitis B vaccine, it was developed to control hepatitis B infection. Hepatitis B is an infectious illness of the liver caused by the hepatitis B virus (HOB) that affects apes, including humans.
The acute illness causes liver inflammation, vomiting, Jaundice, and, rarely, death.Chronic hepatitis B may eventually cause cirrhosis and liver cancer which is a disease with poor response to all but a few current therapies. The infection is preventable by vaccination which is through injecting of recombinant hepatitis B vaccine which contains one of the viral envelope proteins which is hepatitis B surface antigen (Habeas) produced by yeast cell where the genetic code for Habeas has been inserted. By this vaccine injection given, it gives protection against the hepatitis B virus where the immune system provides immunity to hepatitis B infection.Giving vaccination to the indicted person or undetected person especially old age adult NAS not only reduced the risk of infection, but has also led to marked reduction in liver cancer. Besides, the study of molecular biology also developed various techniques which contribute to agricultural sector nowadays.
Through this technique, it capable of improving modern agriculture in producing nutrient-efficient crops which enhancing higher crop productivity. Examples of recombinant DNA technologies which developed for agricultural improvements are golden rice and insect-resistant crops.Golden rice is a recombinant variety of rice that has been engineered to express the enzymes responsible for Гџ-carotene biosynthesize which has high vitamin A content. This variety of rice holds substantial promise for reducing the incidence of vitamin A deficiency in the world’s population who are facing dietary deficiency in their daily food intake which causes blindness, growth retardation and also susceptible to being infected by diseases. While, for the recombinant insect resistant rope has been widely used in agriculture where a bacterium has been applied to crops as an insect-control strategy.
The bacterium was Bacillus trustworthiness that naturally produces a protein (Bet toxin) with insecticidal properties which has insecticide action that is capable of to kill insects. Recently, transgenic plants have been developed by expressing a recombinant form of the bacterial protein, which may effectively control some insect predators that are able to improve agricultural sector. So, through the study of molecular biology by the development of recombinant DNA technology, there are lot of contribution that it can makes on lath and agricultural. Recombinant DNA of human insulin Insulin is a hormone that regulates the amount of glucose (sugar) in the blood and is required for the body to function normally. Insulin is produced by cells in the pancreas, called the islets of Lanterns. These cells continuously release a small amount of insulin into the body, but they release surges of the hormone in response to a rise in the blood glucose level.
Unfortunately, the body of diabetes person could not controlling glucose level in the blood since their body unable to synthesis insulin which causes harmful effect to the person.So that, biosynthesize insulin was developed in order to control sugar level in diabetes suffers body. However, more than half a century of treating diabetics by using animal insulin, but the advance of recombinant DNA technologies produce human insulin by inserting gene of insulin at suitable vector of Escherichia coli. Figure 1: The step of human insulin production Insulin is a protein hormone composed of two chains of amino acids: an “A” chain and a “B” chain linked together by two disulfide bonds.The “A” chain is composed of 21 amino acids and the “B” chain of 30 amino acids, each arranged in a uniquely ordered sequence. Proteins are made by translating the genetic information which is carried in a cell’s genes.
Scientists synthesized in the laboratory genes for the two insulin “A” and “B” chains. This was accomplished by chemically linking together small pieces of DNA sequence and then Joining them in a specific manner to form complete genes.Once the genes were synthesized, they were stitched into circular DNA strands called “plasmids” using special enzymes to perform the molecular surgery.
Plasmids are rings of DNA which are found within the cell. The newly constructed plasmids containing the transplanted genetic material were introduced onto a benign E. Coli bacterial strain. Once inside the bacteria, the genes were “switched-on” by the bacteria to translate the code into either the “A” chain or the “B” chain proteins found in insulin.The process is the same as that used by bacteria to produce its own proteins. When the cells produced sufficient amounts of the “A” and “B” chains, they were harvested to isolate these proteins from the bacteria and purify it. The two chains were then combined chemically in the laboratory to form the complete Insulin molecule which is identical to that produced by the human body. So, the biosynthesize human insulin can be used to diabetes suffer in order to help them controlling glucose level in the blood.
Besides, there is also a few technique was existed in diabetes treatment which involved recombinant DNA technology is Insulin analog. According to Keeper (2001), constructing of insulin analog by changing the structure of native protein improves the properties which can be a great approach of producing synthetic insulin thus helping for diabetes treatment as well. Recombinant DNA hepatitis B vaccine Hepatitis B virus is one of at least three hepatitis viruses causing a systemic infection tit pathological changes in the liver.Infection is transmitted to susceptible persons through close contact with the blood, or other body fluids of chronic infectious carriers or persons suffering acute infection. Because of the urgent need for hepatitis B vaccine, particularly for persons who are at increased risk of acquiring infection, the requirements for hepatitis B vaccine prepared. Since it has been shown that the separated viral coat proteins containing hepatitis B surface antigen lead to the production of protective antibody, it is now possible to use as vaccines.However, the reparation of such vaccines from human viral antigens not grown in cell culture, but obtained from the plasma of infected persons-namely, from persistent carriers of hepatitis B antigens-represents an entirely new approach in vaccine production. A new recombinant DNA hepatitis B vaccine containing hepatitis B surface antigen (Habeas) produced in genetically engineered yeast cells which is Chromosomes crevasses.
Hepatitis B virus DNA (HOB) sequences coding for Habeas obtained from cloned HOB DNA, or, in one case, a partially synthetic sequence, were incorporated into various recombinant plasmids designed tort expression in yeast.Habeas was released by mechanical disruption of yeast cells and was purified by a variety of methods applied sequentially including (a) immunological affinity, (b) hydrophobic interaction, (c) ion exchange, and (d) gel exclusion chromatography. Copying and rate sedimentation centrifugation were also employed. The vaccine was prepared by adsorbing the purified Habeas onto aluminum hydroxide AH(OH)3, the yeast product was shown to be capable of eliciting an anti-Hobs response in laboratory animals.
According to world health organization (1985), chimpanzees minimized withal least woo of the experimental vaccines were shown to resist a live Hepatitis B virus challenge of a thousand infectious doses. The monogenic potency of the yeast product in animals was similar to that of vaccines proved the effective of the hepatitis B vaccine against infectious of Hepatitis B disease. According to Abraham et al. (1999), the fear about these vaccines was the risk of transmission of blood-borne illnesses since the plasma was obtained from affected individuals.This fear seems to be unfounded, where the plasma-derived vaccine having a commendable safety record. These result large amounts of antigen to be produced by recombinant Hepatitis B vaccine prepared from genetically engineered yeast cells.
Figure below showed the overall process of producing HOB vaccine in order to prevent the Hepatitis B infection. Figure 3: The process of producing recombinant Hepatitis B vaccine Recombinant Golden rice: Rice is the major staple food for hundreds of millions of people especially Asian, African and American Latin.The edible part of rice grains consist of the endosperm, filled with starch granules and protein which important to our body. However, it lacks several essential nutrients for the maintenance of health such as Гџ-carotene which is precursor of Vitamin A. Thus reliance on rice as a primary food staple contributes to vitamin A deficiencies, a serious health problem such as blindness and susceptibility to disease that leading to an increased incidence of premature death of small children.So that, there is a way to defeat Vitamin A deficiency in rice which can be achieved through molecular biology technique by introducing the Гџ-carotene biosynthesize pathway into rice endosperm by genetic engineering that we called as “recombinant golden rice”.
According to Miller, HI (2009), recombinant DNA technology applied to agriculture which is has contribute significantly to public the add ion to genes that allow rice to synthesize beta-carotene (the precursor of vitamin A) in its edible endosperm. Golden rice was designed to produce Гџ-carotene, a precursor of vitamin A in the edible part of rice.The rice plant can naturally produce Гџ-carotene in its leaves, where it is involved in photosynthesis.
However, the plant does not normally produce the pigment in the endosperm, where photosynthesis does not occur. So, the formation of Golden rice which introducing p- carotene synthesis of rice endosperm was involved a few steps where the genes that eve golden rice its ability to make Гџ-carotene in its endosperm (interior of the kernel) come from daffodil of Narcissus pseudoscience’s and a bacterium called Erwin redroot.The pathway of Гџ-carotene biosynthesize pathway into rice endosperm by genetic recombinant The pathway toward Гџ-carotene formation required several enzymes which are epiphyte syntheses (sys) and logotype Гџ-cycles (Гџ-LLC) both from Narcissus pseudoscience’s and a bacterial epiphyte decelerates (CTR), under the control of the endosperm-specific gluten promoter. These genes along with promoters (segments of DNA that activate genes) are inserted to plasmids (small oops of DNA) that occur inside a species of bacterium known as Crematorium tendencies.These acrobatic are then added to a Petri dish containing rice endosperm. As they “infect” the endosperm, they also transfer the genes that encode the instructions for making Гџ-carotene. The transgenic rice plants we crossed with strains of rice that are grown locally and are suited to a particular region’s climate and growing condition in order to produce golden rice which has vitamin A content. Through this recombinant of DNA technology, it has potential to make contributions to human health and could enhance the quality of agricultural of reducing nutrient rich-rice.
Recombinant insect resistant crops: Molecular biology technique was used in agricultural sector which is called plant genetic engineering which offers opportunities for the creation of insect-resistant plants by insertion and expression in plant of anthropogenic proteins. An approaches to obtain such plant has been explored which involves the use of delta- indention coding sequences originating from the bacterium Bacillus thirstiness.This approach has being devoted in order to obtaining plants of different species expressing such genes and snowing resistance to insect pests that is increasing crop reduce efficiency. According to Joaquin et. Al (1998), losses due to pests and diseases have been estimated at 37% of the agricultural production world-wide, with 13% due to insects. So, it is necessary to obtain insect-resistant crops which are more friendly agriculture that are able to decrease the percentage of plant losses due to insects.
B. Hurriedness is a gram positive bacterium commonly used in biological pesticide that produces crystal protein (pertinacious inclusions) called d-antitoxins during suppuration. Most B. thirstiness strains produce several crystalline proteins (Cry routines) that have insecticide action.
The d-antitoxins are solubility in the insect midget and are activated by gut proteases that cleave the protein into a smaller polypeptide, the toxin. This toxin binds to the surface of epithelial cells in the midget, inducing lesions that destroy the cells and lead to the death of the insect.According to Lemmas and Peggy (2008), spores and crystalline insecticidal proteins produced by B. thirstiness have been used to control insect pests since the sass and are often applied as liquid sprays.
The approach consisting in the transfer and expression of B. Hurriedness toxin-encoding genes into plants has attracted much attention. Indeed, such a system allows the entire plant to be protected, especially against insects such as borers that infect plant parts by spraying the crops with the liquid sprays. According to Peck et al. 1987) the Belgian company Plant Genetic Systems (now part of Brayer Crosspiece) was the first company in 1985 to develop genetically modified crops (tobacco) with insect tolerance by expressing cry genes from B.
thirstiness. Nowadays, this approach has been used widely in agricultural sector on many crops pieces such as maize, rice, cotton against insects. Figure below show the proven of recombinant insect resistant crops, where the crops that has been sprayed by liquid spray contains of crystalline insecticidal proteins produced by B. thirstiness would not damaged by insects.Figure 2: B. thirstiness toxins present in peanut leaves protect it from extensive damage caused by insect lesser cornstalk borer larvae.
Many studies are under way in order to identify new insecticidal products. One strategy is to screen new sources for potential insecticidal proteins in a random fashion. B. Hurriedness produces a protein that can protect crops against lepidopterist insects such as the black cutworm (Gratis epsilon), a corn pest.
However, Streptomycin cultures secrete a cholesterol oxides active against the boll weevil (Monotonous grand’s).These proteins are highly toxic (within the same range as B. thirstiness toxins) and could be new interesting sources for engineering resistance. According to Joaquin et. Al (1998), because of the bacterial origin of these genes, synthetic genes may have to be constructed to reach high enough levels of expression in order to produce insect rope resistance.
Like all other techniques of molecular biology, recombinant DNA technology has also influenced human life by developing products which are of great benefit to the human body such insulin and hepatitis B vaccine.