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Lab 13 - Protein Synthesis and Genetic Engineering

Lab 13 - Protein Synthesis and Genetic Engineering - Today...

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Unformatted text preview: Today Today Last Full Lab!!! Exam 5 Next Week Turn in Chula Vista Nature Center Turn worksheets and Experiment Report worksheets Handing back: Lab 12 Exam 4 Scenario: Sickle Cell Disease Scenario: Sickle Cell Inheritance Sickle A person can be a person carrier or have the disease disease NOT sex-linked Most prevalent in Most areas with malaria carrying mosquitoes carrying Sickle Cell Disease Sickle Caused by a point Caused mutation on the Hemoglobin Beta Gene (HBB) Gene Results in structurally Results abnormal Hemoglobin HbS HbS Results in periods of Results pain, ranging from minor to major minor Sickle Cells Sickle Red Blood Cells become sickled (flattened) Red after releasing oxygen (can block the blood vessels due to their irregular shape) vessels What’s up with E. coli What’s Bacteria - many strains Bacteria The dangerous stuff is usually found in The livestock digestive tract livestock Sloppy slaughter and messy farms lead to Sloppy contamination of meat contamination Also can be found in sprouts, lettuce, Also unpasteurized milk unpasteurized Once infected, bacteria from feces is Once spread through inadequate hygeine spread What does E. coli do to people? What Most cases Bloody diarrhea and abdominal cramps Fever Resolved in 5-10 days Hemolytic uremic syndrome - destruction of Hemolytic RBCs and Kidney failure RBCs Long term consequences - abnormal kidney Long function, high blood pressure, seizures, blindness, paralysis, bowel surgery/removal blindness, Some cases Laboratory 13 Laboratory Protein Synthesis and Protein Genetic Engineering Genetic Schedule Schedule Start with Investigations 1 and 2 I will go to each group and assist with will investigation 3 investigation 2 Types of Nucleic Acid: Types RNA & DNA Differ in a few essential ways: DNA replication DNA DNA replication DNA Helicase unwinds a Helicase portion of the strand portion DNA polymerase DNA binds and begins replicating in the 3’ to 5’ direction of the original strand original DNA ligase I binds DNA together pieces in the lagging strand (5’ to 3’) 3’) DNA Transcription DNA Protein Synthesis Protein Process where Process DNA encodes for the production of amino acids and proteins. proteins. 1. 2. Occurs in two Occurs steps steps Transcription Translation Transcription Transcription RNA polymerase uses one RNA side of the DNA molecule to synthesize messenger RNA (mRNA) (mRNA) mRNA then migrates to the mRNA cytoplasm, where it is spliced (non-coding sequences deleted) sequences Translation Translation The ribosome binds to the mRNA at the The start codon (AUG) start codon = a 3 nucleotide sequence that specifies a particular amino acid or teminal signal particular The ribosome moves along the mRNA The strand from codon to codon, which codes for amino acids for tRNA molecules with the anticodon bind to tRNA the mRNA codon with an attached amino acid acid Translation Translation These amino acids’s are These then joined together to form a polypeptide chain form The Central Dogma The The CENTRAL DOGMA OF MOLECULAR BIOLOGY The CENTRAL states that information in cells flows from DNA to RNA to protein. to DNA Transcription RNA Translation Like DNA replication, transcription uses the specificity of base pairing to keep the correct order of bases Protein http://chineseculture.about.com/library/symbol/blcc_color.htm Reading the genetic code Reading mRNA - AUGUACUCCGCUAAUUAG Split into 3-letter CODONS Split AUG-UAC-UCC-GCU-AAU-UAG Each codon codes for an amino acid Each (abbr.) building block of a protein (fig 9-2) (abbr.) Full proteins have “start” and “stop” so Full ribosomes know where to begin and end ribosomes reading reading Protein refresher Protein Enzymes are Proteins What if there’s a point mutation in an What enzyme? Is there a particular site on that molecule Is where a change could be disastrous? where Investigation 1 Protein synthesis Investigation Work in pairs FOLLOW DIRECTIONS! Use puzzles to construct the single strand Use of DNA as Figure 9-1. of Transcribe mRNA from the DNA strand. Translate mRNA into proper protein using Translate genetic code table (figure 9-2). genetic Remember DNA – Thymine RNA – Uracil Put puzzle pieces back when finished Investigation 2 Through gene cloning (genetic Through engineering) you give a foreign gene to another species. This species in turn will make the protein coded for by the gene through its protein synthesis mechanisms. through Plasmid Plasmids are circular double stranded DNA molecules that are separate from the chromosomal DNA and usually occur in bacteria. Transformation 1. Use enzymes to cut DNA and Plasmid into desired pieces - specific cuts! 2. Cuts have “sticky ends” open single strands 3. Desired strand of DNA put into plasmid, gets inserted into bacterial VECTOR 4. Not all bacteria gets transformed Investigation 2 Genetic engineering Investigation Work in pairs. A paper simulation to open a plasmid and paper insert a human gene for insulin. insert Choose one of two restriction enzymes. Choose Only one leaves intact both the human Only gene and the antibiotic resistance gene gene in plasmid. in Investigation 3 Transformation of a foreign gene foreign Group of 3 (two groups per bench) Transform a plasmid with the genes for Transform ampicillin resistance and luminescence luminescence found in fireflies to bacteria. found Sterile Technique - For your results and Sterile your health! your Practice with micro-pipettes! Bacteria Transformation Bacteria Investigation 3 Transformation of a foreign gene foreign Control E. coli - Wild Type Not resistant to Ampicillin (AMP) Does not have the gene for bio-luminescence T E. coli - Use this for transformation We will transform them to have a gene for We AMP resistance and one to make them glow in the dark. in Why the AMP resistance? Investigation 3 Transformation of a foreign gene foreign 3 plates per group - LABEL with Group # plates and Section Start time! AMP NO AMP AMP C C T 1 blue line 2 red lines 2 red lines Investigation 3 Transformation of a foreign gene foreign Growth expectations? NO AMP AMP *Stack in groups of six upside down AMP C C T Investigation 3 Transformation of a foreign gene foreign Growth expectations? NO AMP AMP *Stack in groups of six upside down AMP C C T Growth - WT Investigation 3 Transformation of a foreign gene foreign Growth expectations? NO AMP AMP *Stack in groups of six upside down AMP C C T Growth - WT No growth WT not AMP resistant Investigation 3 Transformation of a foreign gene foreign Growth expectations? NO AMP AMP *Stack in groups of six upside down AMP C C T Growth - WT No growth WT not AMP resistant Growth Transformed (any unsuccessful transformations will not grow!) ...
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