biomolecular principles: nucleic acids and dna mohamed bingabr, ph.d. associate professor department...
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Biomolecular Principles:Nucleic Acids and DNA
Mohamed Bingabr, Ph.D.Associate Professor
Department of Engineering and Physics University of Central Oklahoma
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Deoxyribonucleic acid (DNA)DNA Contains all the instructions for producing every molecule currently in the cell (protein).
Human DNA extended 9 cm
storage of genetic information.
Origin of most diseases
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DNA Structure
Nucleotides Nucleic Acids DNA
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NucleotideProvide a mechanism for storage of genetic information.
Adenine Base (A)
Thymine Base (T)
Guanine Base (G)Cytosine Base (C)
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Nucleotides Bonding (1)
The phosphate linked to carbon 5 is coupled to carbon 3 through the hydroxyl (53’).
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Nucleotides Bonding (2)Hydrogen bonding between bases.
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Deoxyribonucleic acid (DNA)
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DNA Fingerprinting (1)Technique: Restriction fragment length polymorphism (RFLP)
1)Restriction enzymes cut the long DNA strands into small fragments. Enzyme EcoRI cuts the DNA at every site that contains the sequence GAATTC.
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DNA Fingerprinting (2)2) Sort the fragment according to size using gel
electrophoresis
3) Press the gel against filter paper to create a replica of the gel.
4) Soak the filter paper in a solution of radioactive DNA probe.
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DNA Fingerprinting (3)1) Radioactive DNA will hybridize with the DNA
samples.
2) Radioactive bands can be photographed with x-ray film, making a picture of the DNA fingerprint.
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Chromosomes
• DNA is folded and packaged into chromosomes to fit inside the nucleus.
• DNA is packaged with proteins called histones to form a material called chromatin.
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Chromosomes• Human cell contain 46 chromosomes arranged in 23
pairs.• The paired chromosomes are called homologous
chromosomes: one is inherited from each parent.• Genes are located on chromosomes.
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Transcription and TranslationHow is the sequence of nucleotide bases in a gene used to synthesize a protein?
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DNA Replication Machinery (1)• Direction of replication 5’ to 3’• DNA Helicase: unwinds the double helix• Single-Stranded DNA Binding: keep DNA from winding
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DNA Replication Machinery (2)• DNA polymerase: synthesis the DNA strand• DNA ligase: join the Okazaki fragments• Topoisomerase: prevents the twisting of the chromosome
Lack of one protein prevent polymerization
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DNA Transcription of a Gene
Promoter codon Termination codon
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mRNA Translation: Protein Synthesis
The organelles ribosomes is a machine that is composed of proteins and package of nuclec acids called ribosomal RNAs
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Genetic Code (1)• Genetic Code: The sequence in which the A, C, G, and T organic bases appear on a strand.
• Gene: determines the instruction for linking amino acids to form protein.
• Every three bases is a code (triplet) for one amino acid.
• GAA is the code for the amino acid Glutamate
• AUG is the start codon and the code for the amino acid methionine
• 3 billion organic bases encode 30,000 human genes
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Genetic Code (2)
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Control of Gene Expression
• All human cells contain the same genetic information
• Cells express only a fraction of the genes
• Different types of cells express different genes
• Example: Only certain cells of the pancreas express
the gene for insulin.
How is gene expression controlled in individual cells?
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Regulation of Gene expression (1)
1. DNA Level: Transcription factors can bind to the promotor regions to enhance or inhibit transcription (steroid hormone)
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Regulation of Gene expression (2)
2. RNA Level: Transcription of unstable mRNA that degraded
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Regulation of Gene expression (3)
3. Protein Level: Protein can be inactivated until they are needed.
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Application of Gene Expression• Block expression of specific gene to study its function.• Gene Therapy: block gene from excessive production of protein.
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Recombinant DNA Technology• Techniques to transfer genetic information from one organism to another.
• Molecular cloning: making a copy of a gene
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Cloning of DNA Using Bacteria• Bacteria used to clone
foreign segments of DNA
• Same enzyme used to cut the plasmid and the foreign DNA.
• Transformation by mechanical or chemical technique
• Transfection: Inserting plasmid to animal cell.
• Transduction: Inserting gene to animal cell by viral method.
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Applications of Recombinant DNA 1
Genetically modified organisms-Golden rice: produce provitamin A and iron-More resistance to environmental damage and viral infection.
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Applications of Recombinant DNA 2Animal Models of Human DiseasesAlter the DNA to produce similar human diseases- to study the efficacy of potential therapeutics to human disease.- to study the function of unknown gene or protein
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Gene Therapy and Viral VectorsRetrovirus are single-strand RNA viruses that infect animal cells and use the host transcription and translation machinery to produce viral proteins (HIV).
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DNA Disorder
InterGenetic Inc in Oklahoma: OncoVue Breast Cancer Risk Test
http://www.intergenetics.com/intergenetics/index.html
Disorder Mutation Chromosome22q11.2 deletion syndrome D 22qAngelman syndrome DCP 15Down syndrome C 21Hemophilia P XColor blindness P X
P - Point mutation, or any insertion/deletion entirely inside one gene D - Deletion of a gene or genes C - Whole chromosome extra, missing, or both - see chromosomal aberrations T - Trinucleotide repeat disorders - gene is extended in length
Currently about 4,000 genetic disorders are known, with more being discovered.
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Cellular Physiological LifestyleAnimation of Inner Cell Life with Music: http://www.studiodaily.com/main/searchlist/6850.html
Animation of Inner Cell Life with Explanation: http://multimedia.mcb.harvard.edu/media.html
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Homework (due March 3)
There are different type of genetic disease. Write a paper (2 - 3 pages) about a disease that is due to one defected gene.The paper should talk about:- Symptoms of the disease-The defected gene and its function-The protein produced by the gene.- The cause of the defected gene (inherited or viral)-Possible cure for the disease-Other interesting things about the disease