Organic Chemistry Lab I
Organic Chemistry Lab I CHEM 2011
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This 17 page Class Notes was uploaded by Columbus Kerluke on Sunday October 11, 2015. The Class Notes belongs to CHEM 2011 at East Tennessee State University taught by Staff in Fall. Since its upload, it has received 10 views. For similar materials see /class/221422/chem-2011-east-tennessee-state-university in Chemistry at East Tennessee State University.
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Date Created: 10/11/15
PowerPoint Lecture Presentation for Concepts of Genetics Ninth Edition Klug Cummings Spencer Palladino Chapter 13 Recombinant DNA Technology and Gene Cloning Lectures by David Kass with contributions from John C Osterman Copyright 2009 Pearson Education inc Chapter Contents 131 Recombinant DNA Technology Combines Several Laboratory Techniques 132 Restriction Enzymes Cut DNA at Specific Recognition Sequences 133 Vectors Carry DNA Molecules to Be Cloned 134 DNA Was First Cloned in Prokaryotic Cells 135 Yeast Cells Are Used as Eukaryotic Hosts for Cloning Continued Chapter Contents 136 137 138 139 1310 1311 Plant and Animal Cells Can Be Used as Host Cells for Cloning The Polymerase Chain Reaction Makes DNA Copies without Host Cells Recombinant Libraries Are Collections of Cloned Sequences Specific Clones Can Be Recovered from a Library Cloned Sequences Can Be Analyzed in Several Ways DNA Sequencing Is the Ultimate Way to Characterize a Clone 131 Recombinant DNA Technology Combines Several Laboratory Techniques Recombinant DNA refers to the joining of DNA molecules usually from different biological sources that are not found together in nature The basic procedure for producing recombinant DNA involves generating specific DNA fragments using restriction enzymes joining these fragments with a vector transferring the recombinant DNA molecule to a host cell to produce many copies that can be recovered from the host cell The recovered copies of a recombinant DNA molecule are referred to as clones and can be used to study the structure and orientation of the DNA Recombinant DNA technology is used to isolate replicate and analyze genes 132 Restriction Enzymes Cut DNA at Specific Recognition Sequences A restriction enzyme binds to DNA at a specific recognition sequence and cleaves the DNA to produce restriction fragments Figure 131 and Figure 132 1 3 1 e r u w4 F Copyright 2009 Pearson Education Inc Figure 132 w m w Staphylococcus aureus 3A Bali Blevibaclerium albidum Haemophilus aegypticus Alul Adhobacter Iuteus Thermus aquaticus Bacillus amyloliqueiaciens H AGT T i i i A TTi F in Haemaphilus in uenzae Rd AATTg i G i Escherichia cali H cleavage sequence Enzyme Recognition Cleavage pattern Source Most recognition sequences are palindromic and restriction enzymes often cleave these sequences in an offset manner Figure 132 Copyright 2009 Pearson Education Inc Copyright 2009 Pearson Education Inc Figure 132 w m w Staphylococcus aureus 3A Bali Blevibaclerium albidum Haemophilus aegypticus Alul Adhobacter Iuteus Thermus aquaticus Bacillus amyloliqueiaciens H AGT T i i i A TTi F in Haemaphilus in uenzae Rd AATTg i G i Escherichia cali H cleavage sequence Enzyme Recognition Cleavage pattern Source DNA ligase joins restriction fragments covalently to produce intact DNA molecules Figure 133 Cleavage with EcaRl 5 39l39l39lquot l 3 Jc TTA A Fragments with complementary tails lt 0 m4 395 5 Copyright 2009 Pearson Education inc Cleavage with EcoRI AAT TCIE 339 3 I 393 539 Annealing allows recombinant DNA molecules to form by complementary base pairingThe two strands are not covalently bonded as indicated by shaded gaps 7 3r I r DNA llgase seals the gaps covalently bonding the two strands Figure 133 133 Vectors Carry DNA Molecules to Be Cloned Copyright 2009 Pearson Education Inc
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