medicinalplantbiotechnology_AND_STUDY_OF_DNA[1].pptx

breif about dna

1. MEDICINAL PLANT BIOTECHNOLOGY md.karishma ., M. Pharm Dept. of Pharmacognosy, au college of pharmaceutical sciences . GENETIC AND MOLECULAR BIOLOGY ,STUDY OF DNA

2. CONTENTS Introduction Historical Perspective• Prospects for Development Application in Pharmacy and Allied fields Genetic and Molecular Biology Study of DNA .

3. INTRODUCTION : Biotechnology is the controlled use of biological agents, such as micro- organisms or cellular components, for beneficial use. Plant Biotechnology describes a precise process in which scientific techniques are used to develop useful and beneficial plants. This includes genetic modification, tissue culture, and molecular breeding to develop plants with desirable traits such as higher yield, disease resistance, and improved nutritional content. Plant biotechnology plays a crucial role in food security, sustainable agriculture, and environmental conservation HISTORY PERSPECTIVES Matthias Schlieden and Theodor Schwan proposed the concept of Cell Theory in the 19 century. Gottlieb Haberlandt, a German bolarist proposed the PatieIn 1981, scientist at Ohio university proposed the first Transgenic Genes from the other animals into mice. In 1980, Genetic Engineering was first used.

4. PROSPECTS FOR DEVELOPMENT OF BIOTECHNOLOGY (OBJECTIVES) 70- 80% of people worldwide believe & turn to traditional herbal medicines. The global demand for herbal medicine is growing gradually Various technologies- adopted for enhancing bioactive molecules in medicinal plants. Biotechnological tools are important for the multiplication and genetic enhancement of medicinal plants. Molecular biology, enzymology, and fermentation technology of plant cell cultures- these systems may become a viable source of important secondary metabolites. DNA manipulation is resulting in relatively large amounts of desired compounds produced by plants infected with an engineered virus. Combinatorial biosynthetic strategies are expected to utilized for important classes of natural products, including alkaloids, terpenoids, and flavonoids. Several genes from different Taxus species are responsible for steps in biosynthesis. It is building a basis for today's combinatorial biosynthesis.

5. PROSPECTS FOR DEVELOPMENT OF BIOTECHNOLOGY AS SOURCE OF MEDICINAL AGENTS 1. Enhanced Production of Existing Medicinal Compounds Plant Tissue Culture: Growing plant cells, tissues, or organs in a sterile, controlled environment (in vitro). Techniques include micropropagation and cell suspension cultures. This allow for the mass production of plant-based pharmaceuticals under controlled conditions. This can ensure a consistent supply of high-quality raw materials, independent of environmental factors. Example: Large-scale production of Catharanthus roseus (Madagascar periwinkle) for its anti-cancer alkaloids. Vinblastine: Used to treat Hodgkin's lymphoma, non-small cell lung cancer, and other cancers. Vincristine: Used to treat leukemia , lymphoma, and other cancers.

6. Elicitation: Plant cell cultures can be treated with elicitors (biotic or abiotic stress factors) to enhance the production of secondary metabolites, which often include the desired medicinal compounds. Example: Using jasmonic acid to enhance the accumulation of Taxol (paclitaxel) in Taxus (yew tree) species cell cultures. Taxol is a crucial chemotherapy drug used to treat various cancers, including ovarian, breast, and lung cancer. Metabolic Engineering: Genetic engineering can be employed to modify the metabolic pathways in plants or plant cell cultures to increase the yield of specific medicinal compounds or to produce novel derivatives with improved therapeutic properties. This could involve upregulating key enzymes in the biosynthetic pathway or introducing new genes. 2. Production of Novel Medicinal Agents Plant-Made Pharmaceuticals (PMPs): Plants genetically engineered to produce therapeutic proteins, vaccines, and other high-value pharmaceutical compounds.

7. APPLICATION OF BIOTECHNOLOGY IN PHARMACY AND ALLIED FIELD Pharmaceutical biotechnology is a relatively new and growing field in which the principles of biotechnology are applied to the development of drugs. A majority of therapeutic drugs in the current market are bio formulations, such as antibodies, nucleic acid products, and vaccines. Production of Antibodies Plants now have potential as a virtually unlimited source of mAbs, referred to by some as 'plantibodies'. Tobacco plants have been used extensively for antibody expression systems, other plants have been used including potatoes, soybeans, alfalfa, rice, and corn. Production of Vaccines The plant species to be used for the production and delivery of an oral vaccine. Corn, is a good candidate for vaccine production for Animals.

8. In humans, particularly infants,the plant of choice is the banana. Cereals and other edible plants are more advantageous than tobacco for vaccine production. The wide variety of other therapeutic agents also have been derived. Eg: Hepatitis – Interferon α ; Anemia - Erythropietin GENETIC & MOLECULAR BIOLOGY APPLIED TO PHARMACOGNOSY Genetics and Molecular biology are field of biology studies the structure and function of genes at a molecular level and thus employs methods of both molecular biology and genetics. The study of chromosomes and gene expression of an organism can give insight into heredity, genetic variation, and mutations. Some of the major applications are,

9. Cultivar Identification: PCR related methods DNA barcoding technology has enormous potential for cultivar identification. It was reported that APAPD and MARMS methods identified five kinds of Panax Ginseng- related species. Resource Protection The information on genetic diversity could guide the protection and development of medicinal resources, especially the rare and endangered ones. Cistanche Deserticola and Cistanche Tubulasa , two endangered medical plants, provided evidence for the protection of wild resources. Formation mechanism of medicinal materials with good quality The quality of medicinal materials is highly affected by their genetic basis and ecological environment. Helpful for Molecular breeding and cultivation of medicinal materials

10. Production of active compounds Transgenic technology has been successfully used to obtain transgenic medicinal plants Which have either disease, insect, drought, and salinity resistance, or have higher production of active compounds. The production of tanshinone was obtained from hairy roots and suspension cells of Salvia miltiorrhiza. STUDY OF DNA : Deoxyribonucleic acid (abbreviated DNA) is the molecule that carries genetic information for the development and functioning of an organism. DNA, or deoxyribonucleic acid, is the hereditary material in humans and almost all other organisms. DNA ,abbreviation of deoxyribonucleic acid , organic chemical of complex  molecular  structure that is found in all  prokaryotic  and  eukaryotic   cells  and in many  viruses .

11. History of DNA The structure of DNA was interpreted only after many types of experimental evidence and theoretical considerations were combined. The crucial evidence was obtained by X-ray crystallography. Some chemical substances, when they are isolated and purified, can be made to form crystals. The positions of atoms in a crystalline substance can be inferred from the pattern of diffraction of X-rays passed through it. Even today, however, this is not an easy task when the substance is of enormous molecular weight. the attempt to characterize DNA would have been impossible without the crystallographs prepared by the English chemist Rosalind Franklin .

12. Franklin’s work, in turn, depended on the success of the English biophysicist Maurice Wilkins , who prepared a sample containing very uniformly oriented DNA Fibers . These DNA preparations provided samples for diffraction that were far better than previous ones. X-Ray Crystallography Revealed the Basic Helical Structure of the DNA Molecule The positions of atoms in a purified chemical substance can be inferred by the pattern of diffraction of X-rays passed through it, although the task requires tremendous skill.

13. CHARGAFF’S RULE As his first step in this search, Chargaff set out to see whether there were any differences in DNA among different  species . After developing a new paper chromatography method for separating and identifying small amounts of organic material, Chargaff reached two major conclusions (Chargaff, 1950) . (a) First, he noted that the nucleotide composition of DNA varies among species (b) the amount of adenine (A) is usually similar to the amount of thymine (T), and the amount of guanine (G) usually approximates the amount of cytosine (C). the total amount of purines (A + G) and the total amount of pyrimidines (C + T) are usually nearly equal. (This second major conclusion is now known as "Chargaff's rule.")

14. Four key features define DNA structure - Four features summarize the molecular architecture of the DNA molecule: It is a double-stranded helix. It has a uniform diameter. It is right-handed (that is, it twists to the right, as do the threads on most screws). It is antiparallel (the two strands run in opposite directions). Structure of DNA

15. WATSON AND CRICK DESCRIBED THE DOUBLE HELIX The solution to the puzzle of the structure of DNA was accelerated by model building : the assembly of three-dimensional representations of possible molecular structures using known relative molecular dimensions and known bond angles. This technique, originally exploited in structural studies by the American chemist Linus Pauling, was used by the English physicist Francis Crick and the American geneticist James D. Watson, then both at the Cavendish Laboratory of Cambridge University. Watson and Crick attempted to combine all that had been learned so far about DNA structure into a single coherent model.

16. The crystallographers’ results convinced Watson and Crick that the DNA molecule is helical (cylindrically spiral) and provided the values of certain distances within the helix. The results of density measurements and previous model building suggested that there are two polynucleotide chains in the molecule. Modelling studies had also led to the conclusion that the two chains in DNA run in opposite directions—that is, that they are antiparallel . Crick and Watson built several large models. Late in February of 1953, they built a model out of tin that established the general structure of DNA. This structure explained all the known chemical properties of DNA, and it opened the door to understanding its biological functions. There have been minor amendments to that first published structure, but its principal features remain unchanged.

17. The band represent the two phos [hate chains Pairs of bases form horizontal connection between the chains Two chains run in opposite direction

18. The sugar-phosphate “backbones” of the polynucleotide chains coil around the outside of the helix, and the nitrogenous bases point toward the centre, The two chains are held together by hydrogen bonding between specifically paired bases. Consistent with Chargaff’s rule, adenine (A) pairs with thymine (T) by forming two hydrogen bonds; and guanine (G) pairs with cytosine (C) by forming three hydrogen bonds. Every base pair consists of one purine (A or G) and one pyrimidine (T or C). This pattern is known as “ C omplementary base pairing ”.

19. Pairs of complementary bases form hydrogen bonds that hold the two strands of the DNA double helix together. Each phosphate group links the 3’carbon of one sugar to the 5’ carbon of the next sugar along the backbone COMPLEMENTARY BASE PAIRING

20. DNA is a double-stranded helix, with the two strands connected by hydrogen bonds. A bases are always paired with Ts, and Cs are always paired with Gs, which is consistent with and accounts for Chargaff's rule. Most DNA double helices are right-handed; that is, if you were to hold your right hand out, with your thumb pointed up and your fingers curled around your thumb, your thumb would represent the axis of the helix and your fingers would represent the sugar-phosphate backbone. Only one type of DNA, called Z-DNA, is left-handed. The DNA double helix is anti-parallel, which means that the 5' end of one strand is paired with the 3' end of its complementary strand (and vice versa).Nucleotides are linked to each other by their phosphate groups, which bind the 3' end of one sugar to the 5' end of the next sugar.

21. Not only are the DNA base pairs connected via hydrogen bonding, but the outer edges of the nitrogen-containing bases are exposed and available for potential hydrogen bonding as well. These hydrogen bonds provide easy access to the DNA for other molecules, including the proteins that play vital roles in the  replication  and expression of DNA. The double helical structure of DNA is essential to its function The genetic material performs four important functions, and the DNA structure proposed by Watson and Crick was elegantly suited to three of them. The genetic material stores an organism’s genetic information . The genetic material is susceptible to mutation . The genetic material is precisely replicated in the cell division cycle. The genetic material is expressed as the phenotype .

22. DNA replication  is the  biological process  of producing two identical replicas of DNA from one original  DNA  molecule.  DNA replication occurs in all  living organisms  acting as the most essential part of  biological inheritance . This is essential for cell division during the growth and repair of damaged tissues, while it also ensures that each of the new cells receives its own copy of the DNA .  The cell possesses the distinctive division property, which makes DNA replication essential. DNA REPLICATION

23. Three modes of DNA replication appeared possible --- The prediction that the DNA molecule contains the information needed for its own replication was demonstrated by the work of Arthur Kornberg, then at Washington University in St. Louis. He showed that DNA can be synthesized in a test - The tube contains just three substances: The substrates, deoxyribonucleoside triphosphates dATP , dCTP , dGTP , and dTTP The enzyme DNA polymerase DNA, which serves as a template to guide the incoming nucleotides

24. There were three possible patterns that could result in complementary base pairing during DNA replication: Semiconservative replication , in which each parent strand serves as a template for a new strand, and the two new DNAs each have one old and one new strand Conservative replication , in which the original double helix serves as a template, but does not contribute to, a new double helix Dispersive replication , in which fragments of the original DNA molecules serve as templates for assembling two

25. DNA RECOMBINANT TECHNOLOGY The process involves the introduction of a foreign piece of DNA structure into the genome which contains our gene of interest. This gene which is introduced is the recombinant gene and the technique is called Recombinant DNA technology. Tools Of Recombinant DNA Technology The enzymes which include the restriction enzymes help to cut, the polymerases- help to synthesize and the ligases- help to bind. The restriction enzymes used in recombinant DNA technology play a major role in determining the location at which the desired gene is inserted into the vector genome. Process of Recombinant DNA Technology Step- 1 . Isolation of Genetic Material The first and the initial step in Recombinant DNA technology is to isolate the desired DNA in its pure form i.e. free from other macromolecules.

26. Step- 2 . Cutting the gene at the recognition sites. The restriction enzymes play a major role in determining the location at which the desired gene is inserted into the vector genome. These reactions are called ‘restriction enzyme digestions’. Step- 3 . Amplifying the gene copies through Polymerase chain reaction (PCR). It is a process to amplify a single copy of DNA into thousands to millions of copies once the proper gene of interest has been cut using restriction enzymes. Step- 4. Ligation of DNA Molecules . In this step of Ligation, the joining of the two pieces – a cut fragment of DNA and the vector together with the help of the enzyme DNA ligase. Step- 5 . Insertion of Recombinant DNA Into Host . In this step, the recombinant DNA is introduced into a recipient host cell. This process is termed as Transformation. Once the recombinant DNA is inserted into the host cell, it gets multiplied and is expressed in the form of the manufactured protein under optimal conditions.

27. Application of Recombinant DNA Technology Gene Therapy – It is used as an attempt to correct the gene defects which give rise to heredity diseases. Clinical diagnosis – ELISA is an example where the application of recombinant Recombinant DNA technology is widely used in Agriculture to produce genetically- modified organisms such as Flavr Savr tomatoes, golden rice rich in proteins, and Bt- cotton to protect the plant against ball worms and a lot more. Recombinant DNA technology enables develop vaccines by cloning the gene used for protective antigen protein. Viral vaccines, through this technology, for example, Herpes, Influenza, Hepatitis, and Foot and Mouth Disease In Industry- production of chemical compounds of commercial importance, improvement of existing fermentation processes, and production of proteins from wastes, also used for the production of Insulin.

28. Purpose Tool/Technique Example 1. Authentication DNA Barcoding ID of Panax ginseng vs Eleutherococcus 2. Adulteration Detection PCR, RAPD Detect Aristolochia in herbal mix 3. Biodiversity Conservation DNA Markers (RAPD, AFLP) Track Taxus baccata populations 4. Chemotaxonomy Phylogenetic Analysis Link Solanaceae plants with alkaloids 5. Quality Control DNA Fingerprinting Standardize Withania somnifera batches 6. Molecular Breeding Marker-Assisted Selection Improve Catharanthus roseus vinblastine Purpose of Studying DNA in Pharmacognosy

29. 1.https://www.nature.com/scitable/topicpage/discovery-of-dna-structure-and-function-watson-397/ 2.https://elearning.raghunathpurcollege.ac.in/files/2BD2BC4915966180050.pdf 3.https://www.biologyonline.com/dictionary/protein-synthesis 4.https://samples.jblearning.com/0763740632/40632_ch08_rev151_188.pdf 5. https://www.slideshare.net/slideshow/introduction-to-plant-biotechnology-durgashree-diwakar/253572600 6. https://www.khanacademy.org/science/hs-bio/x230b3ff252126bb6:gene-expression-and-regulation/x230b3ff252126bb6:dna-structure-and-replication/a/dna-replication 7. https://openstax.org/books/biology/pages/14-introduction REFERENCES