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Biology Project Overview

Biology Project Overview. Comparisons of GAPC Gene Organization in Plants. GAPC Gene. Codes for the enzyme: Glyceraldehyde-3-Phosphate Dehydrogenase (Cytosolic) Gene is found in the cell nucleus Member of a Gene Family

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Biology Project Overview

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  1. Biology Project Overview Comparisons of GAPC Gene Organization in Plants

  2. GAPC Gene • Codes for the enzyme: Glyceraldehyde-3-Phosphate Dehydrogenase (Cytosolic) • Gene is found in the cell nucleus • Member of a Gene Family • 4 genes in the thale cress plant Arabidopsis, a member of the mustard plant family • GAPC and GAPC-2 code for enzymes that act in the cell cytoplasm • GAPCP-1 and GAPCP-2 code for enzymes that act in the chloroplasts

  3. GAPC Gene Product Glyceraldehyde-3-Phosphate Dehydrogenase is the catalyst • Enzyme (gene product) works in the cell cytoplasm • Reaction: • NADH is an electron carrier that donates electrons to the Mitochondrial Electron Transport Chain for production of ATP • ATP (Adenosine Triphosphate) provides energy for most endergonic biological reactions NAD+ (Nicotine Adenine Dinucleotide) accepts one proton and two electrons to become NADH

  4. Using Polymerase Chain Reaction (PCR) • PCR is a highly accurate and efficient way of copying (amplifying) a specific region of DNA for study • The process involves specific primers that attach to the end of the target sequence and serve as starting points for elongation • Heating and cooling cycles provide multiple rounds of PCR where the template number doubles with each round • Taq polymerase, a heat-stable enzyme that catalyzes DNA synthesis, is used to produce the DNA copies

  5. Using Polymerase Chain Reaction (PCR) • Starting with a single copy of a DNA sequence, over 1 million copies can be generated with about 30 rounds of PCR • In the Biology Research Project, students will amplify the nuclear gene for glyceraldehyde phosphate dehydrogenase (GAPC) from a variety of plants, with a “nested” procedure using two sequential sets of primers to enhance specificity

  6. Using Polymerase Chain Reaction (PCR) Inner Arrows show position of primers for PCR Round 2 Outer Arrows show position of primers for PCR Round 1 Sizes of DNA Products can be determined by agarose gel electrophoresis GAPC gene PCR product is missing one intron (interruption) found in the other three genes

  7. ROUND 2 ROUND 1 Positive Control (genomic) Positive Control (genomic) Positive Control (plasmid) Positive Control (plasmid) Negative Control Negative Control Size Markers Size Markers Oregano Oregano Parsley Parsley Comparing Round 1 and Round 2 Results

  8. Join the Biology Research Project if You: • Are interested in applying molecular biology techniques • Polymerase Chain Reaction • Agarose Gel Electrophoresis • Would like to explore evolutionary relationships • Sizing DNA Fragments • Bioinformatics of Sequence Data

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