BIOTECHNOLOGY ISSUES IN SCIENCE AND TECHNOLOGY [C3T1G3]
BIOTECHNOLOGY ISSUES IN SCIENCE AND TECHNOLOGY [C3T1G3] GISAT 113
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This 9 page Class Notes was uploaded by Antonia Lehner on Saturday September 26, 2015. The Class Notes belongs to GISAT 113 at James Madison University taught by Stephanie Batchelet in Fall. Since its upload, it has received 47 views. For similar materials see /class/214150/gisat-113-james-madison-university in General Education Isat at James Madison University.
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Date Created: 09/26/15
GISAT 113 Issues in Living Systems Concept list Unit ll Phenotype v Genotype o Phenotype Characteristic of an organism It is the result of what the genes are in the organism o Genotype the genetic content of the organism It is the cause ofthe phenotype Alleles working definition 0 Two different versions ofthe same gene 0 Each individual has a unique set ofalleles RFLP analysis steps uses and analysis 0 RLFP Restriction Fragment Length Polymorphisms o The basis for DNA fingerprinting Use restriction enzymes to cut DNA at speci c sites 0 Example EcoR cuts at 5 GGATCC3 Alleles with different sequences will have different numbers and locations of RE sites9 different size fragments Steps of RFLP 1 Collect blood tissue etc 2 Cleavage by restriction enzyme 9 different size pieces 3 Separate DNA based on size 9 electrophoresis Genomes size content structure 0 Genome all the genes within an organism o EVERY CELL contains the whole genome 0 Not every gene is used in every cell 0 The type of the cell is defined by the genes they useproteins they make 0 Chromosome colored body unit of genetic material section of the genome 0 Each organism has a set number HUMANS 44 nonsex chromosomes 0 2 copies of 22 2 sex 9 diploid or 2n BACTERIA 1 copy of 12 chromosomes 9 haploid or 1n 0 DNA condensation how it s done reasons for it and why it s important 0 Reasons for it Size must t in nucleus ofthe cell Organization Each new cell must get an ENTIRE genome not some ortoo much 0 Why it s important Transcription when an mRNA copy is made from DNA becomes a polypeptide via translation Euchromatin NOT tightly packed DNA 9 transcription CAN occur Heterochromatin IS tightly packed 9 transcription CAN NOT occur o How it s done Strand of DNA charge is looped around octamers of histone proteins charge These histone loops Nucleosomes are then packed even tighter into zig zags via Linker Histones Radial Loops DNA is looped around proteins that are anchored to the nuclear membrane 0 Variability in looping results in different levels of condensation The nal product is a chromosome 0 Chromatin the DNAProtein complex that results from condensation 0 Active v dividing cells 0 Active Cells keep their DNA loosely packed so they can make mRNA via transcription and ultimately proteins via translation 0 Dividing Cells have very compact DNA to allow for sorting chromosomes Transcription stops Eukaryotic Cell Cycle events and checkpoints o Interphase 1 G1 Mother cell has n chromosomes 2 sets of n2 each 2 DNA Synthesis S Chromosome replication produces n pairs of sister chromotids 0 Each chromosome is copied by DNA Polymerase 0 After replication the DNA condenses so it s easier to sort 3 GZ Replication is completed Cell prepares to divide Replicated chromosomes condense in preparation for M phase Mitosis and Cytokinesis o M Phase 4 Mitosis chromosomes separate 5 Cytokinesis 2 cells are formed from 1 each containing n chromosomes Checkpoint Proteins prevent a cell from progressing through the cell cycle Their role is to check the integrity ofthe genome and prevent a cell from progressing past a certain point in the cell cycle ifa genetic abnormality is detectedPREVENTS CANCEROUS CELLS FROM MULTIPLYING Checkpoint Locations 1 G1 checkpoint 2 G2 checkpoint 3 M checkpoint Homologous chromosomes v sister chromotids o Homologous Chromosomes which are the two different copies ofthe same chromosome that dipliod organisms like humans inherit one from each parent 0 Sister Chromotids identical copies ofa chromosome connected by a centromere Mitosis the division ofthe nucleus into 2 nuclei 0 Function growth and development 0 Asexual reproduction o Wound healing 0 Development 0 Stages and ploidy 2n9 4n 92h 1 Prophase Sister Chromotids condense and spindle starts to form Nuclear membrane begins to disassociate into vesicles 2 Prometaphase Nuclear membrane has completely dissociated into vesicles and the spindle is fully formed Sister chromotids attach to spindle via kinetochore microtubules 3 Metaphase sister chromotids align along the metaphase plate a plane halfway between the poles 4 Anaphase sister chromotids separate and individual chromosomes move toward poles as kinetochore microtubules shorten Polar microtubules lengthen and push poles apart 5 Telophase and cytokinesis chromosomes decondense and nuclear membranes reform Cleavage furrow separates the 2 cells 0 Nondisiunction ERROR in sorting and distributing chromosomes Both chromosomes of a pair are sent to one cell and the other cell gets none 0 Consequences of Mitotic nondisiunction Aneuploidy for a single cell Aneuploidy too many or too few chromosomes 0 What happens next depends on the integrity of the checkpoints 1 Cell unable to repair errors 9 Cell Suicide apoptosis 2 OR cell may become cancerous uncontrolled growth 0 Mechanics of separating chr motor proteins 0 Spindle made ofmicrotubules made oftubulin Forces Generated o tubulin addedremoved 0 motor proteins pull Cohesion to microtubules and between sisters is critical When chromosomes missegregate you get a cell with too little or too few chromosomes big trouble Cancer 2 cause of death for Americans 0 Causes 0 Aneuploidy missegregation of chromosomes during mitosis o Mutations in tumor suppressor or onconogenes Mutagens carcinogens Chemicals or other things that induce DNA mutations Random Mutations o Oncogenes tumor suppressors 1 u Oncogenes when mutated function increased or changed they allow for unchecked cell cycle Tumor Suppressor Genes when mutated function lost policing of cell growth or DNA damage stops 0 Characteristics of Cancer Uncontrolled mitosis Chromosomal instability Mutations in genes lots of replication lots of errors Enlarged nuclei always dividing Resistant to apoptosis No contact inhibition will overcrowd a tissue No anchorage dependence can move throughout the body metastasis Reduced need for growth factors no longer need encouragement from other cells to grow cell immortality lncreased angiogenesis blood vessels feed demand of rapidly growing cells 0 Difficulties in finding a cure The enemy is your own cells how do you kill just the cancer cells and not the whole person Every cancer is different so different treatments are required 0 cell type 0 stage of the disease 0 type ofmutations in the cell 0 Chemotherapy poison to kill ALL dividing cells Meiosis 0 Goal Separation of homologous and later sister chromosomes within 2n cells into in gametes egg sperm for sexual reproduction DNA is replicated once Cell is split twice Each offspring has a unique combination of Mom and Dad 0 Genetic Variation Random Fertilization Homologous recombination Result four haploid cells called gametes Sexual Reproduction offspring receive 12 of the genes from each parent 0 Meiosis vs Asexual Reproduction Meiosis generation of 1n gametes eggs and sperm to come together to be a 2n baby Asexual Reproduction clonal division into two independent cells bacteria mitochiondria chloroplasts o Ploidy 2n 4n 2n 1n 0 The Human Lifecycle 1 Diploid Adult 2n 0 Meiosis Haploid gamete 1n spermegg Diploid zygote 2n 1 sperm 1 egg Mitosis 4 Development into diploid adult 2n 46 Stages of Meiosis 1 G1 S GZ priorto meiosis 2 Meiosis I rst the homologous pairs are separated into two cells 0 Bivalent tetrad homologous pairs of sister chromotids associate with each other lying side by side formed via synapsis 3 Meiosis N then the sister chromatids are separated into two cells DN 2 Chances for Nondisiunction in Meiosis 1 Nondisjunction of homologous chromosomes in meiosis l 2 Nondisjunction of sister chromotids on meiosis ll Conseguences of meiotic nondisiunction Aneuploid organism Down Syndrome is one of very few VIABLE human aneuploidies extra copy of chromosome 21 trisomy 21 0 Most aneuploidies are fatal leading cause of miscarriages Maternal age and nondisiunction oogenesis and spermatogenesis 0 Eggs sit in lnterphase l of Meiosis for decades 0 After puberty eggs are released to nish meiosis one at a time o Breakdown of sister chromatid cohesion occurs over time 9 Nondisjunctionaneuploidy 0 Male sperm are produced throughout a man s life no waiting around Meiosis vs Mitosis o Meiosis o Daughter cells different than parent cell 0 4 daughter cells 0 23 chromosomes each 0 Mitosis o Daughter cells same as parent cell 0 2 daughter cells 0 23 pairs of chromosomes each Property Mitosis Meiosis Property Mitosis Meiosis DNA replication During interphase During interphase Divisions One Two Cross over events NA Tetrads in Prophase l Daughter cells genetic composition Two diploid identical to parent cell Four haploid different from parent cell and each other Role in animal body Produces cells for growth and tissue repair Produces gametes Sex determination in different species XYI etc o Humans o 46 chromosomes total 0 O O O 22 sets of 2 sutosomal chromosomes 2 sex chromosomes Females are XX 9 all X eggs Males are XY 9 either X or Y sperm Gregor Mendel s experiments scoring phenotypes to deduce the presence ofalleles Homozygous v Heterozygous o Homozygote individual with identical alleles o Heterozygote individual with differentalleles when talking about recessive alleles that cause disease these individuals are called carfers Using Punnet Sguares in genetics Inheritance atterns dominantrecessive incom lete dominance sexlinked o Incomplete Dominance red ower white ower pink flower 0 SexLinked o Allele is present on a sex chromosome 0 Males are XY females are XX 0 For Xlinked recessive alleles females have a backup 0 Examples color blindness hemophilia Drawing and analyzing simple pedigreessee genetics worksheet Genetic screening for genetic disorders Cystic fibrosis 0 Genetic disorder defect inherited o Defect in a passive membrane chloride transporter mucus is too thick 0 CFTR sttic Eibrosis Iransmembrane conductance Begulator 0 People with errors bad alleles in the CFTR gene don t make Jnctional CFTR protei 0 Over 1000 faulty alleles identi ed 0 Most common error is AF508 a recessive allele PCR How it works denaturing annealing extending What it s used for What s needed in a reaction primers template DNA Polymerase dNTPs Structure of DNA Negatively charged sugarphosphate backbone Base pairing rules and hydrogen bonding o A T o C G 0 Hydrogen bonds broken by DNA Helicase 0 Broken by heat in PCR DNA replication 1 Separation of Strands by DNA helicase strand 2 Polymerization ofthe new a DNA Polymerase 3 Result Replication of both strands of DNA S miconservativ model DNA replication produces dsDNA with 1 parental and one new stran elicase unwinds parental DNA at replication fork Topoisomerase helps prepare DNA for helicase nicks the backbone ofthe DNA in order to relieve supercoiling ssBP ep bubble open a lcmplalc DNA Pnlymerase I and DNA Polymemsel iepiaeesme R NApiimeisWiih DNA DNA Poikmmsemean anaeme pHmEYS and eiengaie Leading and lagging strands 1 DNA pniymerzse can any add nueiemmes inihe 339 end nnhe eh leading and lag strand LIGASE 2 DNA pniymerzse can any add In a Free n9 slrzndgt primers PRIMAS 39 adds NNA pnmeis eempiememaiy RNA Wjumsma and nueieeiieesiegemei 7 Leading strand 6 5mm RNA piimar lt1 Duemiun ulfork f Okalak quot35mm mavemem f Lagging srarid 7 mm minimquot Favemal DNA dumex Okazaki iagmems and hgase Diiemien eipeiymenzaiien Puipuse Ennis eEi 1 Nnei cuiiecied 3 DNA vephcated immme bad cupyWiH eemamme muiaiien paiches eimuiam eeiis z Pleareading DNA Peiymeiase Hi can chew back and iepiaee 3 Mismatch repay MuiH L s sysiem We mistake Peiymeiase backiiacks and Wes mistake immediate y Mismaich YEpaW Nueieeme emsmn Yepaw mus1 cummun o UVdamage TT links 0 modified bases 0 missing bases 0 crosslinks 0 Direct repair of modified base rare 0 Double stranded break repair Transcription Genes encode proteins with functions 0 Genes don t DO things create phenotypes o PROTEINS DO THINGS 0 Only genes that are expressed transcribed and translated become proteins Gene 39 Overview DNA strand 339 5 Template can be the template A c c A A A c c e A c T top or bottom strand strand 3 strand RNA polymerase mRNA 5 Cotton Ribosome TRANSLATION L l Protein Amino acid Open reading frame bases encoding a protein has a specified start and stop Intergenic region bases do not encode a protein 1 spacers between ORFs 2 regulatory regions sites for protein and enzyme binding 3 pseudogenes Iost ORFs have a lot of the characteristics of a gene but have multiple stops along their length 4 97 of human genome Template v nontemplate DNA strands Sequence of events initiation elongation termination Enzymes involved Promoter architecture Eukaryotes v Prokaryotes Monocistronic v polycistronic messages operons Introns and exons mRNA processingsteps and functions Errors Translation Features of mRNA RBS start stop codons
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