deficient‚ recombinant strain. The LEU2 gene is a linear fragment that does not contain an Autonomous Replication Sequence‚ so it could not replicate on its own and needed to be integrated by homologous recombination. The TRP1 gene was a circular plasmid that contained an ARS‚ which allowed for it to act as an extra chromosome in the gene. The objective was to insert a “wild gene” and replace the defective genes and then grow them on a medium that does not contain TRP1 or LEU2 to prove that the genes
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AP Bio - Modern Genetics Protein Synthesis • Start with primer • New strand is 5’ to 3’ • TATA Box - TTAATTAA • RNA Polymerase - Reads and matches bases (One recipe; only reads leading strand) • Single strand produced; mRNA • Now produced pre-mRNA (You need exon‚ not intron) • Introns create spaces‚ need ligase to connect exons to make true mRNA. • Adds a poly A tail (on 3’ side) and 5’ (prime) cap (on 5’ side) used for defense • Leaves through pore to ribosome. • Messenger RNA will attach to
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|This is the cells power plant; it is responsible for the production of energy for the | |(Eukaryotic) |cell. | |Plasmid |Plasmids are an extra-chromosomal DNA that contains genes with special properties that | |(Prokaryotic)
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isolating cells à disrupting lipid membranes with detergents à destroying proteins with phenol or proteases à degrading RNAs with RNase à leaving DNA at the end 2. Crude isolation of plasmid vector DNA is accomplished by an alkaline lysis procedure or by boiling cells which removes bacterial chromosomal DNA from plasmid DNA. 3. To get purer DNA from either (1) or (2)‚ crude DNA is a) Fractionated on a CsCl2 gradient b) Precipitated with ethanol c) Poured over a resin column that specifically binds
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the DNA. 3. When a restriction enzyme cuts out a portion of DNA‚ it will sometimes leave a sticky end. If two fragments of DNA are cut by the same restriction enzyme‚ the sticky ends can join together‚ forming a recombinant DNA strand. 4. Cut plasmid and eukaryotic DNA with the same restriction enzyme… Mix fragments and allow them to match… Ligate… Transform into bacteria… Identify desired clone. 5. a.) The addition of a fluorescent agent can help identify a recombinant cell with a gene of interest
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Momordica charantia The phytochemical prospection of the fresh and dried leaves extracts‚ fruits‚ seeds‚ and bark of the plant contains resins‚ alkaloids‚ flavonoids‚ glycosides‚ steroids‚ terpenes‚ cardiac glycoside‚ saponins having various medicinal importance viz. anti-HIV‚ anti-plasmodial‚ shigellocidal‚ anti-diarrheal‚ anti-septic‚ anti-bacterial‚ anti-viral‚ anti-inflammatory‚ anti-microbial‚ hypoglycemic‚ antioxidant‚ analgesic and hepatoprotective properties. PubMed.gov http://www.ncbi
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AP ESSAY ANSWERS: 16-20 1. Information transfer is fundamental to all living organisms. For TWO of the following examples‚ explain in detail‚ how the transfer of information is accomplished. A) The genetic material in one eukaryotic cell is copied and distributed to two identical daughter cells. B) A gene in a eukaryotic cell is transcribed and translated to produce a protein. C) The genetic material from one bacterial cell enters another via transformation‚ transduction or conjugation
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In the first part of this lab‚ E.coli cells were transformed with an R-plasmid carrying a tetracycline resistant gene‚ giving rise to tetracycline resistant E.coli strain. This was accomplished through transformation‚ which allowed E.coli to directly uptake the naked DNA molecule carrying the antibiotic resistant gene (1). However‚ in order to take up the DNA and incorporate them into their genome via recombination‚ cells must be competent (1). Therefore‚ E.coli cells which are not competent under
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1) What are the technical challenges for cloning a mammal organism? Include the following terms in your answer: cell determination‚ cell differentiation‚ and nuclear reprogramming. Cloning is producing a cell line or culture all whose members contain identical copies of a particular nucleotide sequence; an essential element in genetic engineering (Raven et al‚ G-5). Lack of imprinting; genes that are imprinted are expressed differently‚ the original parent in which the gene was taken from is the
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Recombinant DNA 1. Isolate DNA from 2 sources * Bacterial plasmid w/2 genes * ampR(resistance to ampicillin) * lacZ(makes β galactosidase) * Human cell 2. Treat both w/ the same restriction enzyme * Recognition sequence cuts through lacZ gene in plasmid 3. Mix all DNA fragments with plasmids. * H-bonds bt/n bases temporarily hold “sticky ends” together 4. Add DNA ligase: creates permanent covalent bonds 5. Reintroduce plasmid to bacteria via transformation 6. I.D. recombinant cells
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