Chapter_2_-_Basic_Molecular_Biology_2009

Chapter_2_-_Basic_Molecular_Biology_2009 -...

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Basic Molecular Biology 2.1 Nucleic acid structure 2.1.1 The DNA backbone ( key concept: stability of dsDNA ) 2.1.2 The base pairs ( key concept: melting temperature, hybridization - salt / temp. ) 2.1.3 RNA structure ( key concept: instability of ssRNA ) 2.1.4 Nucleic acid synthesis ( key concept: DNA requires primer, dNTPs added to 3’OH ) 2.1.5 Coiling and supercoiling ( key concept: DNA coiling affects DNA migration rates during gel electrophoresis ) 2.2 What is a gene? ( key concept: gene concept is imprecise ) 2.3 Information flow: gene expression ( key concept: critical gene elements required for bioengineering in vitro transcription and translation ) 2.3.1 Transcription 2.3.2 Translation 2.4 Gene structure and organization ( key concept: identification of elements for deciphering mRNAs, CDS in genomic DNA ) 2.4.1 Operons 2.4.2 Exons and Introns
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2.1 Nucleic Acid Structure 2.1.1 The DNA backbone DNA is DNA is inherently stable Repetitive phosphate-sugar backbone 5’ P links one sugar to the 3’ position of the next P, O, C atoms are covalent bonds Degradation of DNA require nucleases that break these covalent bonds Endonucleases (cut internal sites) Exonucleases (nibble ends) Discovery of restriction endonucleases which cut DNA strands at specific positions Discovery of DNA ligases which join DNA DNA strands Availability of restriction enzymes and DNA ligases opened up the field of genetic engineering Figure 2.1 DNA backbone
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2.1 Nucleic Acid Structure 2.1.1 The DNA backbone RNA contains the sugar ribose rather than deoxyribose in DNA RNA is less stable than DNA Greater susceptibility to degrade by ribonucleases and chemical degradation by alkaline conditions Figure 2.2 Nucleic acid sugars
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2.1 Nucleic Acid Structure 2.1.2 The base pairs Besides covalently bonded backbone 2’deoxyribose sugar and phosphate, DNA contains 4 nitrogen-containing bases Larger purines are matched with smaller pyrimidines in hydrogen bonded pairs Figure 2.3 Nucleic acid bases Figure 2.4 Base-paring in DNA One purine matches with a pyrimidine in complementary strand Hydrogen bonding between AT and GC
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2.1 Nucleic Acid Structure 2.1.2 The base pairs Strands are antiparallel, running 5’ (phosophate) to 3’ (hydroxyl) in either direction Convention for the double stranded sequence of positive strand: 5’ AGGCTG 3’ Negative strand 5 ’CAGCCT 3’ is complementary and reversed of positive strand The negative strand has complementary bases (eg. T for A and C for G), and reversed of positive strand Reading the negative strand is from right to left on the bottom strand (in the 5’ to 3’ direction) Box 2.1 Complementary sequences 5’-AGGCTG-3’ 3’-TCCGAC-5
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2.1 Nucleic Acid Structure 2.1.2 The base pairs DNA is easily denatured in the test tube or
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This note was uploaded on 07/10/2010 for the course BIOL 208 taught by Professor Chuong during the Spring '09 term at Waterloo.

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Chapter_2_-_Basic_Molecular_Biology_2009 -...

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