Professional Documents
Culture Documents
Genetic Microbial
Genetic Microbial
Nucleic acids
Microbial Genetics
DNA
RNA
DNA Replication
Gene expression
Regulation of Genes
Mutation
Gene transfer
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Basic Structure
Base
PO42- 5’
4’ 1’ Adenine
Guanine Purine
Sugar Thymine
Cytosine Pyrimidine
3’ 2’ Uracil
Ribose (OH),
Deoxyribose (H)
Nucleoside
Nucleotide
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DNA : Deoxyribonucleic acid
DeoxyriboNucleicAcid
2 nanometers (nm)
1 turn ~ 10 bp = 3.4 nm
minor groove
major groove
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Organization of
DNA in Cells
(Chromosome)
Prokaryote Supercoiled
ÊǹÁÒ¡¨ÐÁÕŠѡɳÐ໹ǧ¡ÅÁ (Closed
circular) 1 ǧ ÍÂÙã¹ Cytoplasm
à¡ÅÕÂǤ٢ͧàʹ DNA ¨ÐÁÕ¡ÒúԴµÑÇã¹ÅѡɳÐ
"Supercolied" Relaxed circle
ÅѡɳСÒèѴµÑǢͧ Chromosome ໹ÃÙ»ÃÒ§
·Õèä Áṹ͹ àÃÕ¡ÇÒ "Nucleoid"
Nucleoid = DNA (~60%) + RNA (mRNA) +
Protein (Nucleoid prot., Transcription factor)
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Eukaryote
Chromosomal DNA ÍÂÙã¹ Nucleus
ÍÂÙã¹ÅѡɳТͧ DNA-protein complex ·Õè
àÃÕ¡ÇÒ "Chromatin"
Protein ·Õèà¡ÒÐÍÂ١Ѻ DNA ÊǹãË ¤×Í Histone
Histones ÁÕ 5 ª¹Ô´: H1, H2A, H2B, H3, H4
DNA ·Õ¾è ѹÃͺ Histone 8 ˹Ç ä´Å ѡɳÐ໹
àÁç´ ·ÕèÁÕàʹ¼Ò¹Èٹ¡ ÅÒ§ ~10 nm àÃÕ¡ÇÒ
"Nucleosome"
5
RNA
RNA ໹ polynucleotide ÊÒÂà´ÕÂè Ç
Êǹ»ÃСͺ¤ÅÒ¢ͧ DNA ¤×Í »ÃСͺ´ÇÂ
RiboNucleicAcid Phosphate group, Ribose áÅÐ Nitrogen bases
RNA µÒ§¨Ò¡ DNA ¤×Í
¹éíÒµÒÅ໹ Ribose
Base Uracil (U) á·¹ Thymine (T)
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tRNA rRNA
tRNA ÁÕ “Anti-codon” «Ö§è ¡ç¤Í× ÅíҴѺ¡ÒÃàÃÕ§µÑǢͧ ໹ͧ¤ç»ÃСͺ¢Í§ Ribosomes «Ö§è ÁÕº·ºÒ·ÊíÒ¤Ñ-ã¹
Nucleotides 3 µÑÇ ·ÕÊè ÒÁÒöà¢Ò¤Ù¡Ñº "Codon" º¹àʹ ¡Ãкǹ¡Òà Translation
mRNA
»ÅÒ´ҹ 3' ¢Í§ tRNA ¨Ðà¡ÒСѺ Amino acid ·Õè
¨íÒà¾ÒÐ
´Ñ§¹Ñ¹é tRNA ¨Ö§à»¹µÑÇàª×Íè ÁÃÐËÇÒ§ÅíҴѺàºÊ·Õè໹ÃËÑʺ¹
àʹ mRNA áÅÐÊÒ¢ͧ Amino acid ÊíÒËÃѺ
¡Ãкǹ¡Òà Translation à¾×èÍãËä´ Protein
Replication Translation
Transcription
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DNA Replication
Replication
Semi-conservative = ¡ÒÃÊѧà¤ÃÒÐË• DNA ãËÁ
â´Â¡Ò÷Õè DNA à´ÔÁ«Öè§à»¹à¡ÅÕÂǤÙÍÂÙ¹Ñé¹ á¡ÊÒÂ
ÍÍ¡ÁÒ໹àʹà´ÕèÂǡ͹ ¨Ò¡¹Ñé¹áµÅÐÊÒ·íÒ
˹ҷÕè໹áÁẺ (Template) ã¹ ¡ÒÃÊѧà¤ÃÒÐË•
polydeoxyribonucleotide ÊÒÂãËÁ¢Öé¹ÁÒ ¼Å·Õèä´
¤×Í DNA ·ÕèÊÃÒ§¢Öé¹ÁÒãËÁ¨Ð»ÃСͺ¢Ö鹨ҡ
polydeoxyribonucleotide ÊͧÊÒ â´ÂÊÒÂ˹Öè§
ÁÒ¨Ò¡ DNA à´ÔÁ ÍÕ¡ÊÒÂ˹Öè§ÁÒ¨Ò¡¡ÒÃÊѧà¤ÃÒÐË•
¢Öé¹ãËÁ
8
Mechanisms of Replicationtion in E. coli
DNA (àʹ¤Ù) ¤ÅÒÂà¡ÅÕÂÇ໹ÊÒÂà´ÕèÂÇ ·Õè oriC
(Helicase, Single-stranded DNA binding
proteins (SSBs), DNA Gyrase)
ÊÃÒ§ RNA primer ÊÒÂÊÑé¹ æ (Primase)
µÍÊÒ polynucleotides ãËÂÒÇ¢Öé¹ â´Â¡ÒÃàµÔÁ
dNTPs (5'3') ·Õè»ÅÒ¢ͧ RNA primer â´Âãª
ÊÒÂà´ÔÁ໹áÁẺ (DNA polymerase III)
DNA polIII ¨ÐÊѧà¤ÃÒÐË• DNA ã¹·ÔÈ·Ò§¨Ò¡ 5'3'
´Ñ§¹Ñé¹ DNA 2 àʹ·ÕèÊà ҧ¢Öé¹ãËÁ¨ÐµÒ§¡Ñ¹
9
RNA synthesis
Transcription Transcription
ÊÇ ¹·ÕèàËÁ×͹ DNA replication 㪠RNA polymerase 㹡ÒÃÊѧà¤ÃÒÐË• RNA ¨Ò¡
ÁÕ¡ÒÃàµÔÁ dNTPs ã¹·ÔÈ·Ò§¨Ò¡ 5’ ä» 3’ DNA
ÊÇ ¹·ÕèµÒ§¨Ò¡ DNA replication Template ¤×Í ÊÒ“Anti-sense”
ãªá¤ºÒ§ÊÇ ¹¢Í§ DNA ໹áÁẺ (template) µíÒá˹§àÃÔÁè µ¹¡Òà Transcription ·ÕèÍÂÙº¹àʹ
㪠RNA polymerase 㹡ÒÃàµÔÁ dNTPs
Template ¤×Í “Promoter region”
¨Ø´ÊÔ¹é ÊØ´¢Í§¡Òà Transcription ¤×Í “Terminator
àÁ×Íè àÊÃç¨ÊÔ¹é ¡Ãкǹ¡Òà ¨Ðä´ single stranded RNA
region”
10
Mechanisms of Transcription
RNA pol ¨Ðà¢Ò ¨Ñº¡Ñº DNA ·ÕèµíÒ á˹§ Promoter
áÅФÅÒÂà¡ÅÕÂÇ DNA ÍÍ¡ áÅÐÊѧà¤ÃÒÐË•ÊÒÂ
RNA ã¹·ÔÈ·Ò§ 5' ä» 3'
àÁ×èÍ RNA pol à¤Å×è͹·Õè件֧ Terminator region
¡ÒÃÊѧà¤ÃÒÐË• RNA ¨ÐËÂشŧ RNA pol áÅÐ RNA
·ÕèÊѧà¤ÃÒÐË•¢Ö鹨ÐËÅØ´ÍÍ¡¨Ò¡¡Ñ¹ áÅÐËÅØ´¨Ò¡
DNA
Protein synthesis
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Translation The major participant in Translation
¡ÒÃá»Å¢ÍÁÙÅ·ÕèºÃèØÍÂÙº¹ mRNA ä»à»¹ÅíҴѺ¢Í§ amino mRNA: ºÃÃ¨Ø Codon (3nt.), µíÒá˹§¨´¨íҢͧ
Ribosome, Start & Stop signals ÊíÒËÃѺ
acid ·Õ¨è Òí à¾ÒÐ (Protein)
¡ÒÃá»ÅÃËÑÊ໹ protein
¡Ãкǹ¡Òà Polymerization ¢Í§ amino acids ä» tRNA: ºÃÃ¨Ø Anti-codon (ÁÕ¤ÇÒÁ¨íÒà¾ÒСѺ codon)
໹ Polypeptide sequence â´ÂÁÕâÁàÅ¡ØŢͧ Ribosome: à绯ԡÔÃÔÂÒ¡ÒÃÊÃÒ§¾Ñ¹¸Ð Peptide
Ribosome áÅÐ tRNA ÁÒà¡ÒСѺ mRNA à¾×èÍ¡ÍãËà¡Ô´ ÃÐËÇÒ§ Amino acid ã¹ÊÒ Protein
¡ÒÃÊѧà¤ÃÒÐË•ÊÒ Polypeptide
12
tRNA Genetic code
13
Elongation Termination
14
Genetic regulation
15
Lactocs (lac) operon Lactocs (lac) operon
RNA Polymerase
RNA Polymerase
Mutation
Mutation ¡ÒÃà»ÅÕè¹á»Å§·Õèà¡Ô´¢Öé¹ã¹ÊÒþѹ¸Ø¡ÃÃÁ ໹¼Å
ãËÁÕ¡ÒÃáÊ´§ÅѡɳзÕèà»ÅÕèÂ¹ä» áÅÐÊÒÁÒö¶ÒÂ
·Í´ÅѡɳйÑé¹ä»ÊÙÅÙ¡ËÅÒ¹ä´
16
Causes of Mutation Types of Mutation
17
Insertion or Deletion (No Framshift)
18
Substitution (Nonsence mutation)
Gene transfer
Transformation
Transduction
Conjugation
19
Transformation
¡ÒÃà¤Å×è͹ÂÒÂÊÒþѹ¸Ø¡ÃÃÁ (DNA) ¨Ò¡áº¤·ÕàÃÕÂà«ÅÅ•
˹֧è ä»ÂѧẤ·ÕàÃÕÂÍÕ¡à«Åŕ˹֧è
à«ÅÅ·ÕèÃѺÊÒþѹ¸Ø¡ÃÃÁµÍ§ÁÕÊÀÒ¾·Ò§ÊÃÕÃÇÔ·ÂÒ·Õè àËÁÒÐÊÁ
àÃÕ¡à«ÅÅ•·ÃÕè ºÑ DNA ¹ÕéÇ Ò "Competent"
à¡Ô´ã¹áº¤·ÕàÃÕª¹Ô´à´ÕÂǡѹ áµµÒ§ÊÒ¾ѹ¸Ø ËÃ×Íà¡Ô´ã¹
Ấ·ÕàÃÕµҧª¹Ô´¡Ñ¹¡çä´
Ấ·ÕàÃÕÂ㹸ÃÃÁªÒµÔ·àÕè ¡Ô´ Transformation ä´§Ò હ
Acinetobacter, Azotobacter , Bacillus ,
Streptococcus, Haemophilus
Transduction
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Conjugation
¡ÒÃà¢Ò¤Ù¡ ѹ¢Í§áº¤·ÕàÃÕ áÅÐÁÕ¡ Òö·ʹÊÒþѹ¸Ø¡ÃÃÁ
(DNA) ãËá¡¡ ѹ â´ÂẤ·ÕàÃÕÂà«ÅÅ˹Ö觨Ð໹¼ÙãË
(Donor) áÅÐÍÕ¡à«ÅÅ˹Öè§à»¹¼ÙÃѺ (Recipient)
à«ÅÅ•¼Ùã ˨ÐÁÕ F plasmid (F+) áÅÐÊÒÁÒöÊÃÒ§
"Sex pilus" à¾×èÍãªã¹¡ÒèѺ¤Ùá ÅÐʧ DNA à¢Òä»ã¹à«ÅÅ
¼Ùà Ѻ (F-) J. Lederberg & E. Tatum (1958)
à¡Ô´ã¹áº¤·ÕàÃÕ¡ÅØÁ·ÕèÊÒÁÒöà¢Ò¤Ù¡Ñ¹ä´
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