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This 5 page Study Guide was uploaded by Lisa Montanez on Saturday April 23, 2016. The Study Guide belongs to 300 at Edinboro University of Pennsylvania taught by Dr. William J. Mackay in Winter 2016. Since its upload, it has received 33 views. For similar materials see Genetics in Biology at Edinboro University of Pennsylvania.
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Date Created: 04/23/16
GENETICS STUDY GUIDE CHAPTERS 20, 21, 22, 24 & 26 INTRODUCTION Biotechnology: an application of biology for commercial purposes o Genetic engineering manipulate genes o Recombinant DNA technology o Transgenic organisms CHAPTER 20: RECOMBINANT DNA TECHNOLOGY Clones o Recombinant clone Chimeric molecule: 2 parts- donor DNA + vector DNA Chimera: greek mythology- creature ½ man ½ beast 4 Steps 1. Isolate DNA a. Donor: genomic b. Vector: plasmid- mini, midi, maxi 2. Cut DNA using restriction enzymes a. Restriction enzyme: endonuclease i. Internal substrate enzyme (nucleic acid), recognizes palindromes in DNA 1. Palindromes: breaks bonds in a restriction site ii. Produced by bacteria iii. Protects bacteria from viruses 3. Join DNA using complimentary base pairs and ligase 4. Amplify DNA- transformation o Cellular Clones By asexual reproduction o Therapeutic Clones: stem cells o Reproductive Clones: stem cells VECTORS: 5 GENERAL TYPES 1. Plasmids a. Accepts 5 kb DNA b. pBR318 c. PUC o Smaller o Altered origin: multi copy o amp (Ampicillin) o Modified lacZ’ operon lacZ’ codes for p-galtetramerpolypeptide with 1173 amino acids, 146 amino acids ar N terminus o Multiple cloning site(MCS), poly linker A 200 base pair invitro DNa sequence that contains many restriction sites DH5 α Restore function to lacZ , special complamentation o Alpha (α) complementation for lacZ lacZ+¿ o X-GAL -- blue indole B−GAL ¿ Blue/white screen to detect donor molecules d. pBluescript 2. Phages: accepts 15 kb donor DNA 3. Viruses (cosmids): accepts 45 kb donor DNA 4. YAC’s: yeast artificial chromosomes: accepts 1 mb donor DNA 5. BAC’s: bacterial artificial chromosomes: accepts 3-6 mb donor DNA λ 45 kb linear virus with 2 life cycles: o Lisogenic o Lytic: arms + donor DNA Concatemer: very large molecule consisting of multiple viroal genomes Confluent cellsbacterial lawn Plaques: clear zone in a bacterial lawn which is the result of a successful viral infection Library: collection of DNA sequences that represent an organism placed into vectors o Genomic library: genome In genome: coding sequence, exon, introns, promotors, regulatory sequences, moderately repetitive DNA (transposable elements, SINES, LINES), highly repetitive DNA o cDNA library: m RNA HYBRIDIZATION SYSTEMS o Southern Blot: DNA to DNA hybridization o Northern Blot: DNA to RNA hybridization o Western Blot: protein- antigen (Ag) produce antibody (Ab) Ag Ab detection system Autoradiography: detects radioactive hybrid nucleic acid molecules DNA Sequencing: a technique used to identify and order DNA molecules which differ in size by 1 bp using electrophoresis or automation o Ribose DNA: 2 OH groups o Deoxyribose: 1 OH group o Dideoxyribose: no OH groups Will stop reaction, reaction termination Polymerase Chain Reaction: (PCR), used to modify DNA 1. Denaturation: dsDNA ssDNA, denature at 95 C 2. Renaturization: with primers, 50-55 C 3. Extension: using DNA polymerase, 70-72 C o PCR has 30 cycles o DNA pictogram 10 -1 microgram 10 -6 o Heat stable DNA polymerase: Taq polymerase withstands 95C Signst Transduction Pathway 1. 1 messenger 2. Receptor 3. Relay protein 4. Effector (enzyme) nd 5. 2 messenger (AMP) 6. Cellular response o 1 messenger is hormones, neurotransmitters, growth factors Yeast- unicellular eukaryote o Cell cycle: 1. Isolate mutants (cell cycle defective mutants) temperature sensitive 2. Group mutants: complementation analysis =300-500 genes a. Regulatory: cyclins- proteins that fluctuate in concentration i. Cell cycle dependent kinase (CDK)- requires cyclin to function - b. Energy: kinase (enzyme)- donates PO gr4ups (from ATP) to a target molecules i. Maturing Promoting Factor (MPF)- complex cyclin + CDK, maturing promoting factor cell cycle CHAPTER 24: CANCER Cancer: progressive disease, cancer genes, uncontrolled cell division Carcinogens: chemical, physical, biological o Carcinogenesis: process of getting cancer 1. Initiation: mutagens transform the cell 2. Promotion: confers the cell growth advantage a. Mutagenic b. Non mutagenic (signal transduction pathways) 3. Progression: metastasis: movement of cancer cells from the primary site to multiple secondary sites Mutagenic 4 Basic Tissue Types 1. Epithelial: carcinoma (90%) skin cancers 2. Connective: sarcomas (bone), leukemia (blood), lymphoma (lymph) 3. Muscle 4. Neurons 3 Cancer Genes 1. Oncogenes (proto-oncogenes) 2. Tumor suppressor genes 3. Tumor susceptibility genes TYPES OF TUMORS: o Benign: will not metastasize, localized o Malignant: will metastasize, invasive tumors, invasion is called metastatic Transformation: the process of turning a regular cell into a cancerous cell o V- gene from virus o C- gene from your body CHARACTERISTICS OF CANCER CELLS o Lose cell density inhibition o Require less nutrients o Secrete large amounts of proteases Digest basal lamina o Secrete growth factors Angiogenesis (vascularization) o Less adhesive (E cm) (extravascular matrix) o Fail to differentiate o Fail to undergo apoptosis Programmed cell death Facilitated by caspases (proteases) Transfection: addition of DNA to mammalian cells GLEEVEC: smart drug, specific noncompetitive inhibitor Proto-oncogenes: normal cellular genes that can be mutated into oncogenes, cellular counterpart of oncogenes o Expressed at lower levels relative to oncogenes o “Regular” version of oncogenes Oncoproteins: products of oncogenes o Protein tyrosine kinesis o Growth factors o G proteins (signal transduction) o Transcription factors o Ras- G protein or relay protein Hormone y receptor k Ras(g protein) RasMEKMAPK Ras, MEK, MAPK are intacellular transductors Serine/Tyrosine (S/T) Kinases TUMOR SUPPRESSOR GENES o Normal active cellular gene that represses cell growth o Inactivation or suppression leads to deregulated growth o Cancers caused resulting from inactivation of tumor suppressor genes are inherited in a dominant fashion o Tumor suppressor genes act in a recessive manner (heterozygosity in non-tumorgenic) o Loss of heterozygosity inactivates the remaining wildtype allele (LOH) LOH: loss of heterozygosity Mutate 1 wildtype allele, lose the chromosome containing the second wildtype allele by nondisjunction P53: master cancer gene (60%) o Tumor suppressor gene o Tetramer Complex genetic structure Complex gene structure/ function o BRAC-1/BRAC-2: genes involved in breast cancer Ex: Colon cancer o Normal colon cells APC: tumor suppressor gene DDC: tumor suppressor gene rasK: oncogene p53: tumor suppressor genemetastatic tumor cells TUMOR SUSCEPTIBILITY GENES o Does not actively cause cancer o Increases incidence of cancer (increases frequency of mutation) o Involved in DNA repair o UV light P53P21Rb1---E2F P53: master cancer gene, TSG, blocks DNA replication, induces apotosis P21: cyclin dependent kinase inhibitor Rb1: retinal blastoma protein E2F: transcription factor that tells the cell to not go into cell cycle o No UV light no P53 no P21 Rb1 is phosphorylated o In a child with tumors, Rb1 is phosphorylated and goes through the cell cycle and cancer cells replicate CHAPTER 26: POPULATION GENETICS Population: group of organisms in a given area Gene pool: total number of alleles in a population Allele frequency: percentage of alleles in a population 1908- Hardy Weinberg st o p= frequwncy of the 1 allele o q= frequency of the second allele o p+q = 1 gene pool, haploid o p +2pq+q = 1 diploid NEED 5 CONDITIONS 1. Large population: minimizes genetic drift a. Genetic Drift: changes to the allele by chance 2. Isolated population: minimizes gene flow a. Gene Flow: movement of genes in and out of a population 3. No net mutations 4. Random mating 5. No natural selection Zygotic Gene Expression: wildtype is expressed over recessive gene Maternal Gene Expression: mother’s genes are always expressed o Phenotype of mutant depends of genotype of mother o If mother has function, offspring will also have function regardless go genotype
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