CHAPTER 5: THE SOURCE OF HEREDITARY

Genetically modified strain of mice Doogie greater memory: modification + insertion of NR2B gene improves functioning of nerve receptors memory and learning.

5.1 Early Developments in Genetics

History: early physicians and scientists investigations, artists sketching guide to anatomy, structure of organs: principle: structure function. Study of Genes + technology: Light microscope, electron microscope, X-ray diffraction, gel electrophoresis better picture of mechanisms of gene action. Cytology and Genetics 2000 years ago: Greek philosopher Aristotle: hereditary: power of males semen Other scientists: female determined characteristics, male gamete set events in motion Other theory: hereditary traits in blood: e.g. bloodline 1831: discovery of nucleus towards understanding structure + function of cells + genes. 1865: Mendel published papers. Knowledge: egg + sperm = zygote. Accepted: factors blended = characteristics of offspring. Mendel didnt know meiosis, structure or location of hereditary material, how genetic code worked many theories about inheritance explained how traits were passed. Interpreted as experiment w/ garden peas. Same time. New techniques lens grinding = better microscopes new branch: Cytology: study of cell formation, structure, function. Aided 1882: Walter Fleming: separation of threads in nucleus = mitosis Same ear Edouard van Benden: sperm and egg cells 2 chromosomes = fertilized egg 4 1887- August Weisman: special division theory: reduction division (meiosis) framework for Mendels work 1900: rediscovered Mendels work. -

5.2 Development of the Chromosomal Theory

1902: American Walter S. Sutton, German Theodor Boveri independently chromosome pairs segregate during meiosis, forming new pairs after. homologous chromosomes supported Mendels explanation of inheritance + paired factors. Today: alleles of a gene, each from each sex cell. Cellular evidence explained + supported union of 2 alleles in offspring, formation of new combos in offspring. Chromosomal behaviour gamete formation explained law of segregation + independent assortment Sutton and Boveri deduced factors (alleles) chromosomes. (46 chromosomes = thousands of traits) Sutton hypothesis: each chromosome genes (on same chromosome: linked genes) Chromosomal Theory Development/refinement of microscope cell biology + genetics. Later: biochemistry + nuclear physics Chromosomal theory of inheritance: 1) chromosomes genes, units of hereditary, alleles w/ specific locus/position 2) paired chromosomes segregate (meiosis). Sex cells #, unlike somatic cells. Each gamete: 1 of 2 Assort independently: chromosome pair no influence on movement of others. E.g. AaBb AB, aB, Ab, ab equal frequency -

5.3 Morgans Experiments and Sex Linkage

American geneticist Thomas Hunt Morgan (1866-1945) gender and inheritance Drosophila Melanogaster 1) reproduces rapidly, mating after leaving egg, 100 eggs each time. Study many in short period, larger number ideal for probability. 2) small many in single culture tube small, solid nutrient. 3) Genders distinguishable. Males smaller, rounded abdomen, dark coloured posterior segment. Females larger, pointed abdomen, pattern of dark bands. Mutations (heritable chance molecular structure of DNA) linked to other traits (supported theory: genes chromosomes). Examining eye color. White eyed male among red-eyed offspring decided mutation traced inheritance of allele of white eyes, White-eyed male x Red-eyed female F1: all red eyes Hybrid x Hybrid F2 Red White all females red, half males white half males red Cytology th 8 chromosomes: females 4 pairs, males 3 pairs: 4 pair, sex, partially homologous: X chromosome + Y chromosome. Conclusion: different genes. Explained: Y didnt eye colour (differences: on part of X not matching Y). = Traits on sex chromosomes = sex-linked traits

X X (pure-breeding, red eyed female. Allele red eye dominant, located X chromosome) x X Y (White R r R eyed male: Y: no allele). Offsprings: Red eyes: females X X , males X Y F1 male x female => F2. Male offsprings sex-linked trait is from mother. Father supplies Y. R r R R R r F2 male X Y and X Y. Females: X X or X X . White-eyed females: female with 1 allele for white eyes x white-eyed male. Females 3 genotypes, males 2: cant be homozygous for X gene only 1 X In humans: recessive allele on X for red-green colourblindness. More males (only 1 X = only 1 recessive allele expressed). E.g. human recessive lethal (trait: both alleles death or severe malformation of offspring) X-linked disorders more frequently in males. I.e. females live longer. Autosomal Dominant Genetic Disorders Controlled by dominant alleles on autosomal chromosomes. E.g. Huntingtons disease: lethal genetic disorder rare dominant allele: breakdown of certain areas of brain. No effective treatment. Males + females: equal frequency Autosomal Recessive disorders Recessive alleles on autosomes: needs 1 recessive allele from each parent. E.g. Cystic Fibrosis: most common lethal genetic disease among Caucasians thick mucus affects lungs and pancreas. Pedigree: symbols identifying males, females, individuals affected by trait, family relationships for genetic inheritance.

R R

5.4 Looking Inside the Chromosome

Deoxyribonucleic acid, DNA only molecule that can replicate cellular reproduction. Contains instructions, transmitting hereditary info. Uniqueness new combos of genes and mutations. E.g. instructions: new cells develop into specialized cells, mature cells replacement and repair of worn cell parts. Info chemical messages, nucleus to cytoplasm. How genes affect expression of traits how DNA regulates production cell protein (major structural/functional components). Searching for the Chemical of Hereditary Early 1940s biologists hypothesis: hereditary material Chromosomes. Chromosomes: equal amounts of proteins amino acids and nucleic acids. Altering sequence = new proteins. Nucleic acids similarities. Phosphate group + five carbon sugar molecule + nitrogenous bases = nucleotide. Polymer/long chain = DNA. At first: scientists: protein component of chromosome genetic material w/ hereditary message, master molecule, directing arrangement of amino acids cytoplasm. Nucleic acids monotonous repetitions of 1 sugar molecule, identical phosphates, 4 nitrogenous bases in all living things. Incorrect. -

5.5. Discovering the Structure of DNA

James Watson child prodigy University of Chicago (15). Studied ornithology (study of birds), and genetics/molecular biology. 1951 Englands Cambridge University, met Francis Crick, physicist. interpret and synthesize experimental data.

Source of data: Cambridge laboratory of Maurice Wilkins: Rosalind Franklin: X-ray diffraction structure of DNA molecule. Technique: photographed DNA molecule clear images w/ helical structure + where phosphate sugars were located. Another clue: Scientists knew: DNA sugars (deoxyribose), phosphate, 4 different nitrogenous bases. They didnt know arrangement. New research in any species: # of adenine molecules = thymine molecules, # guanine molecules = cytosine. I.e. arranged in pairs Watsons background (emerging chemical data) + Cricks background (significance X ray diffraction results) 3d model of DNA molecule. 1953 scientific community. 1969 visually confirmed. Current version: extra info Models: useful tools. E.g. DNA: shows how different atoms interact. X-ray diffraction picture how different chemical bonds interact. Visual devices relationship/interactions of diff parts of molecule. Politics and Science Franklin X ray diffraction technique to view image. Watson + Crick, in England model. American scientist Linus Pauling wanted to study, denied visa - identified communist sympathizer, support of antinuclear movement. -

5.6 Structure of DNA

DNA molecule: nucleotides: deoxyribose sugars, phosphates, nitrogenous bases. Double helix/spiral ladder: sugar + phosphate molecules (backbone), nitrogenous bases (rungs) paired by hydrogen bond (weak forms between + of hydrogen atom and on electronegative nitrogen/oxygen atom at end of another molecule). Pair = complementary base pair. Estimate: 3.5 bill base pairs of DNA, 30 000 genes on 46 chromosomes. Nitrogenous bases w/ double rings, purines (adenine and guanine) always combine w/ nitrogenous bases w/ single rings, pyrimidines (cytosine and thymine). Adenine molecule always + thymine (2 hydrogen bonds), guanine molecule always + cytosine (3 hydrogen bonds)

5.7 Genes that Change Position

Barbara McClintock (American) theory: Jumping Gene Theory (genes can move positions). Old theory until 1980s believed genes + chromosomes = fixed. Experiments w/ Indian corn. Colour variation in kernels hypothesized elements: transposons (specific segments of DNA move along chromosomes). Inserting some genes new position of chromosome inactivates genes affection pigment production. 1940s not well accepted. 1983 Nobel Prize. Recombinant DNA Bacterium inserts genes along circular chromosome. Transposon can move chromosome of bacterium to another. Inserted genes can integrate w/ plasmid (in bacterium) secondary structure, small ring of genetic material (cons. Extra DNA) Some bacteria: conjugation: sexual reproduction genetic material exchanged; 2 cells fuse, plasmids passed. Possible to insert DNA from 1 into another genetic engineering. 1 technique recombinant DNA DNA fragments from 2+ organisms spliced together. DNA from 1 cut at specific sites by restriction enzyme DNA fragments w/ unpaired nitrogenous bases/ sticky ends. Same w/ plasmid DNA. 1 fragment another, bonds - complementary nitrogenous bases. Negative conjugation: disease-causing bacteria evolved genes resistant to antibiotics. Normally: interfere w/ chemical reactions in harmful microbes. Now genes of resistance. -

5.8 Genetic Research and Technologies

Human Genome Project mid 1980s: plans mapping entire genetic makeup of human being. Began U.S.A. Oct 1990 James st Watson (1 of 1 directors) Human genome(complete set of instructions in DNA): 30 000 genes w/ 23 pairs of chromosomes w/ 3 bill pairs of nucleotides DNA. Beginning: only 4500 genes + sequence of nucleotides that made up genes. Completed May 2000: many scientists, diff countries, improvements in sequencing techniques. British biochemist Frederick Sanger DNA sequencing technique for project: pieces of DNA replicated + changed so fragments (ending with nucleotides) detected by laser. Automated equipment exact number of nucleotides in chain. Computer combines data + reconstructs original sequence Quantity of DNA initially: cloning human DNA in single celled organisms. Now: American biochemist Kary Mullins millions of copies of 1 molecule of DNA, few hours Add to list of genes for hereditary disorders (cystic fibrosis, muscular dystrophy, Huntingtons chorea new drugs + genetic therapies Other hand: controversial ethical questions, legal dilemmas, societal problems imputable Huntingtons Chorea Incurable brain disorder strikes prime of life debilitating mental breakdown, eventual death Trapped inside own bodies, unable to communicate 1993: Dr Michael Hayden UBC team isolated gene Now: simple DNA test screen people w/ family history, 98% accuracy potential sufferers or parents who might pass on Another technique: animal modeling: lab mice genetically manipulated like people w/ Huntingtons. Studied how and why disorder brain cells to die prematurely. Goal: treatments alleviate effects Cystic Fibrosis Inherited disorder 1 gene protein CFTR (cystic fibrosis transmembrane conductance regulator): must inherit 2 defective alleles (from each parent). 1/25 of European ancestry carries most common recessive genetic disorder Hospital for Sick Children samples from families w/ more than 1 child suffering Dr Lap-Chee Tsui + team identified gene 1989 mapped 2 modifier genes in animals alter severity of cystic fibrosis. Still investigating impact on treatment. U of T: Dr Christine Bear used cystic fibrosis mouse model possible correct defect delivering normal CFTR protein. Working method to provide protein therapy to correct defect. Field of genetics daily medical practice. Dr. Judith Hall UofT: head of pediatrics: clinical geneticist + experience w/ families affected by cystic fibrosis. Communicates genetic reasons to children + families. Muscular Dystrophy Group genetic disorders: weakening + deterioration of muscles. Some forms because defects on autosomal chromosomes, occurs in males + females. Other forms: sex-linked, males. Emery-Dreifuss muscular dystrophy males, recessive X linked gene. Even female carriers normal dominant allele + recessive defective allele: mild symptoms. Hospital for Sick Children: Dr. Ron Worton: located Duchenne muscular dystrophy gene: 1987. Sex linked form boys age 2-6. Progressive damage in muscles of pelvis, upper arms, legs. Calf muscles enlarge: enzyme creatinine kinase leaks swelling. Age 12: wheelchair. Other forms: both males and females equally: defective genes on autosomes. Some forms: recessive and dominant.

5.9 DNA Fingerprinting

DNA fingerprinting test: Alec Jeffreys (geneticist from University of Leicester. Found: long stretches of DNA molecule are similar in everyone: less than 1% unique.

Particular segments unique sequences of nitrogenous bases no function. Belief: nonsense codes, repeat as a chemical type of stuttering DNA fingerprinting test: DNA isolated from skin, hair, semen, blood vs. DNA from blood sample from suspect. DNA samples cut w/ restriction enzymes (possible to cut at specific points) reproducible DNA fragments. Differences in molecular structure = location and number of cuts that can be made are distinct profile of DNA is unique. Transferred nylon sheet, radioactive marker identifies unique sequence of DNA chain placed onto X-ray film. Black bands appear (where markers attach segments used to establish identity) Print from film, used to compare samples.

5.10 Gene Therapy

3500+ genetic diseases liked to defective genes: e.g. cystic fibrosis, diabetes, hemophilia, Huntingtons chorea, sickle cell anemia Treatments: e.g. diabetics insulin injections can control affliction. No true cures. Medical advances + modified diet + restricted behaviour + environmental adjustments reasonably productive life. Potential of transforming the genetic gene Gene therapy: defective genes replaced with normal genes. 3 strategies: 1. Gene insertion: normal gene position by a virus, or other agent, on chromosome of affected cell (not every cell uses gene) to function in the intended manner. 2. Gene modification: defective gene modified chemically recode genetic message. More delicate + needs greater knowledge of chemical composition of normal + defective genes. 3. Gene surgery: defective gene removed + replaced w/ normal gene. First clinical use: 1990: 4 year old girl treatment inherited disorder of immune system enzyme deficiency insufficient amounts of enzyme ADA (adenosine deaminase). Genetically modified virus carried normal ADA gene immune cells. = cells produced required enzyme. Viruses reproduce DNA into normal body cells, reprogramming production of new viruses. Can be modified beneficial genetic material. Trick: inserting in appropriate place (no unexpected results) 1999: McGill Michel Tremblay, Dr. Brian Kennedy found gene controls product - enzyme tyrosine phosphatase, involved in regulating blood glucose levels. Significant role diabetes mellitus (Type II diabetes) and obesity. lead to gene therapy techniques to control problems. 2000: joint team: South Korea + Canada treatment of juvenile (type I) diabetes in mice gene therapy.