Ch3-120127 - CHEM 350: Introduction to Biological Chemistry...

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Unformatted text preview: CHEM 350: Introduction to Biological Chemistry Brian Lee, Ph.D. [email protected] Office: Neckers 146G or 324 Phone: 453-7186 Hours: 9:30am to 10:30am or by appointment Website: https:/ / Textbook (required, U.S. edition only) Fundamentals of Biochemistry, 3rd Ed., Voet, Voet & Pratt. Study Guide (recommended) Student Companion to Fundamentals of Biochemistry, 3rd Ed. Help Desk Tuesday 6:30 to 7:30 pm in Neckers 218 Thursday 5:00 to 6:00 pm in Neckers 410 Announcements Undergraduate Research Opportunities Research for credit (such as CHEM 396 or CHEM 496) Student worker ($8.00 per hour) ( Undergraduate Assistantships ( McNair Scholars Program ( REACH Awards Competition ( (due Jan 30th) Undergraduate Scholarships (due Jan 31st) Chemistry Majors see Department website Scholarships.pdf Science Majors see College of Science website Summer Research Experiences for Undergraduates (REU) Assignments Read Chapter 4 Amino Acids Chapter 4 Problems • First Midterm Exam, Monday February 6th – Chapters 1 through 5 • Help Desk Tuesday 6:30-7:30pm in Neckers 218 Thursday 5:00-6:00pm in Neckers 410 Recombinant DNA Manipulating DNA to study isolated genes or to control expression of genes Plasmid DNA carrier or vector for cloned DNA Cloned DNA - Amplification - DNA Sequencing - Gene Expression Other vectors: bacteriophage, baculovirus, artificial chromosomes (BAC, YAC) How do we select a successful clone? Cloning site disrupts the lacZ gene in the plasmid. LacZ is beta-galactosidase which normal cleaves lactose On agar plates containing Xgal failed clones turn blue Successful clones produce no LacZ and are white DNA Libraries can be stored in larger vectors: bacteriophage, bacterial artificial chromosome (BAC) or yeast artificial chromosome (YAC) vectors for genomic sequencing. Finding a sequence of interest through in situ hydridization Requires a known sequence to probe the library Libraries can be genomic DNA or complementary DNA cDNA library are reverse transcription copies of mRNA Polymerase Chain Reaction – isolate and amplify DNA 1. Denature 2. Anneal 3. Polymerize Second cycle: Repeat 1. Denature Note: Daughter strands are shorter than the parent strands. The new strands begin and end at the primer sequences. 2. Anneal 3. Polymerize (not shown here) At the end of the second cycle, we have unit length strands define by the location of the 5’ and 3’ primers. Further cycles will amplify these products. The PCR product can then be cloned into a vector for transfection in another organism. Transgenic organisms express proteins from a foreign source. Site Directed Mutagenesis to change genes (and produce new proteins) A synthetic primer can be used to introduce a mutation. The new sequence can alter the amino acid composition of the expressed protein in transgenic cells. ...
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This note was uploaded on 03/26/2012 for the course CHEM 350 taught by Professor Lee during the Spring '08 term at SIU Carbondale.

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