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DNA, Proteins, and Biotechnology Mr. Nagel Meade High School Warm Up DNA Replication • Outline the process of DNA Replication. Include enzymes and their functions. IB Syllabus Statements • 3.5.1 – Compare the structure of RNA and DNA. • 3.5.2 – Outline DNA transcription in terms of the formation of an RNA strand complementary to the DNA strand by RNA polymerase. • 3.5.3 – Describe the genetic code in terms of codons composed of triplets of bases. • 3.5.4 – Explain the process of translation, leading to polypeptide formation. • 3.5.5 – Discuss the relationship between one gene and one polypeptide. http://click4biology.info/c4b/3/Chem3.5.htm Nucleotide – Structure Structure of DNA Sense v. Antisense DNA • Sense: mRNA sequence exists that matches it • Antisense: DNA Compliment (usually copied from) • DNA strand 1 (5’3’): sense • DNA strand 2 (3’ 5’): antisense • mRNA codon: sense • tRNA anticodon: antisense Transcription • http://users.rcn.com/jkimball.ma.ultranet/Bi ologyPages/T/Transcription.html – What are the steps? – Are there any special modifications to the mRNA after creation? Transcription • Moving from DNA to RNA – Create a messenger (mRNA) – Moves from the nucleus to the cytoplasm • Specifically the rough ER (ribosomes) – To be read as ‘blueprints’ for proteins • Post-processing – 5’ Cap; Poly-A tail; Intron removal [eukaryotes] Video Time! • http://www.hhmi.org/biointeractive/dna/ani mations.html Think… • How many different words can you make with one of these LETTERs (ATGC)? • TWO LETTERs (ATGC)? • THREE LETTERs (ATGC)? • FOUR LETTERs (ATGC)? Translation Overview • Initiation • Elongation • Termination Arming the tRNA • Each tRNA is shaped differently – Loop regions • Shape is worked on by different enzymes • Enzyme attaches specific AA to tRNA – Requires ATP • Each AA has one or more tRNA molecules – Degenerate code Translation: How it works • rRNA holds the mRNA and brings it together with tRNA http://users.rcn.com/jkimball.ma.ultranet/BiologyPages/T/Translation.html Transcription/Translation Activity • Transcribe the following: – TAC GCC CGA ATA TAT AAA ATT • Translate the following: – AUG GCC UGA Warm Up Translation • In which direction will translation occur? • What will the next amino acid be in the sequence? • Name two places in a eukaryotic cell where translation occurs. Code Game • There are multiple DNA fragments hidden throughout the room (some with multiple copies). – Find the DNA fragments and record them. – Transcribe the mRNA fragments. – Use the Nagel-codon table to decode the fragments. • Once you do so, determine what the original sequence of DNA said (before fragmentation)! DNA Codes (Hidden) • • • • • 1) 2) 3) 4) 5) TAC TTT TCT TAA TTA CTT TAC AGG GCT GGG TCC CAC TGG CGG AAA ACT Mutations • Insertion or Deletion – Frameshift Mutation • Substitution – Point Mutation • Nonsense Mutation • Silent Mutation Warm Up DNA • Determine the strand of mRNA that is transcribed from the strand below. ATC CAG GTC AAG • List three other molecules apart from mRNA that are needed for transcription. • What would the anticodons of the corresponding four tRNA molecules be? Would they be considered sense or anti-sense? IB Syllabus Statements • • • • • • • • • • • • • 4.4.1 – 4.4.2 – 4.4.3 – 4.4.4 – 4.4.5 – 4.4.6 – 4.4.7 – Outline the use of polymerase chain reaction (PCR) to copy and amplify minute quantities of DNA. State that, in gel electrophoresis, fragments of DNA move in an electric field and are separated according to their size. State that gel electrophoresis of DNA is used in DNA profiling. Describe the application of DNA profiling to determine paternity and also in forensic investigations. Analyse DNA profiles to draw conclusions about paternity or forensic investigations. Outline three outcomes of the sequencing of the complete human genome. State that, when genes are transferred between species, the amino acid sequence of polypeptides translated from them is unchanged because the genetic code is universal. 4.4.8 – Outline a basic technique used for gene transfer involving plasmids, a host cell (bacterium, yeast or other cell), restriction enzymes (endonucleases) and DNA ligase. 4.4.9 – State two examples of the current uses of genetically modified crops or animals. 4.4.10 – Discuss the potential benefits and possible harmful effects of one example of genetic modification. 4.4.11 – Define clone. 4.4.12 – Outline a technique for cloning using differentiated animal cells. 4.4.13 – Discuss the ethical issues of therapeutic cloning in humans. http://click4biology.info/c4b/4/gene4.4.htm DNA Tech: PCR DNA Tech: Restriction Enzymes • Restriction Enzymes – Cut like chemical scissors at various recognized sequences DNA Tech: Gel Electrophoresis • Proteins or DNA travel through a gel based on SIZE and CHARGE. • Virtual Gel Electrophoresis – http://learn.genetics.utah.edu/content/labs/gel/ DNA Profiling Cloning (Genes or Organisms) • Bacteria used for genes – Cut and paste using restriction enzymes • Egg cells with removed nuclei are used for cloning entire organisms – Moral or ethical ramifications? DNA Sequencing • Utilize ddNTPs to halt elongation • Millions of strands of various lengths Microarrays and Gene Therapy • Microarrays are used to detect what genes are activated at any given time • Gene Therapy is when defective genes are removed and replaced by working ones, than reinserted into the host Warm Up DNA Revisited • State the names of the structures listed above. • State the name of the structure above that has an anticodon. • Explain why the process above is called translation. • How is translation terminated? IB Syllabus Statements • • • • • • • • • • • • • 4.4.1 – 4.4.2 – 4.4.3 – 4.4.4 – 4.4.5 – 4.4.6 – 4.4.7 – Outline the use of polymerase chain reaction (PCR) to copy and amplify minute quantities of DNA. State that, in gel electrophoresis, fragments of DNA move in an electric field and are separated according to their size. State that gel electrophoresis of DNA is used in DNA profiling. Describe the application of DNA profiling to determine paternity and also in forensic investigations. Analyse DNA profiles to draw conclusions about paternity or forensic investigations. Outline three outcomes of the sequencing of the complete human genome. State that, when genes are transferred between species, the amino acid sequence of polypeptides translated from them is unchanged because the genetic code is universal. 4.4.8 – Outline a basic technique used for gene transfer involving plasmids, a host cell (bacterium, yeast or other cell), restriction enzymes (endonucleases) and DNA ligase. 4.4.9 – State two examples of the current uses of genetically modified crops or animals. 4.4.10 – Discuss the potential benefits and possible harmful effects of one example of genetic modification. 4.4.11 – Define clone. 4.4.12 – Outline a technique for cloning using differentiated animal cells. 4.4.13 – Discuss the ethical issues of therapeutic cloning in humans. http://click4biology.info/c4b/4/gene4.4.htm DNA Technology Please discuss with a neighbor answers to the following: What is it? PCR Restriction Enzymes Gel Electrophoresis (DNA Profiling) Cloning DNA Sequencing DNA Microarrays How is it used in modern science? Bioethics – Small Group • The Case of Cats and Clones – http://auth.mhhe.com/biosci/genbio/olc_linked content/bioethics_cases/g-bioe-03.htm – Read the case study and then answer the questions in your small groups. Discuss your responses in a respectful manner. – Record your responses on the back of the worksheet. Bioethics – Self Study • Given the advent of biotechnology, there seems to be limitless possibilities (curing genetic disorders, engineering our own offspring, etc). – At what point should a line be drawn? – Should there be a line drawn at all? – Who should establish those guidelines?