‘OUTLINE’

Nucleic acids

Microbial Genetics
DNA
RNA
DNA Replication
Gene expression
Regulation of Genes
Mutation
Gene transfer

Nucleic Acids Nucleotide

Polymer ¢Í§ ¹ÔǤÅÕ âÍä·´• (Nucleotide) »ÃСͺ´ŒÇ 3 ʋǹ ¤×Í
ËÃ×Í Ë¹‹Ç‹͠Nucleotides µ‹Í¡Ñ¹à»š¹ÊÒ 1. Pentose : 5 carbon atoms ÁÕ 2 ª¹Ô´ ¤×Í
Polynucleotide 1.1 Ribose (RNA)
à¡çºáÅж‹Ò·ʹ¢ŒÍÁÙÅ / Åѡɳоѹ¸Ø¡ÃÃÁ 1.2 Deoxyribose (DNA)
ÁÕ 2 ª¹Ô´ ¤×Í 2. Nitrogen bases : 2 ¡Å؋Á
DNA : Deoxyribonucleic acid 2.1 Purine (2 rings)
RNA : Ribonucleic acid 2.2 Pyrimidine (1 ring)
3. Phosphate group

1
 Basic Structure

Base
PO42- 5’

4’ 1’ Adenine
Guanine Purine
Sugar Thymine
Cytosine Pyrimidine
3’ 2’ Uracil
Ribose (OH),
Deoxyribose (H)
Nucleoside
Nucleotide

2
 DNA : Deoxyribonucleic acid
DeoxyriboNucleicAcid

RNA : Ribonucleic acid

2 nanometers (nm)

1 turn ~ 10 bp = 3.4 nm

minor groove
major groove
3
 Organization of
DNA in Cells
(Chromosome)

Prokaryote Supercoiled
ʋǹÁÒ¡¨ÐÁÕŠѡɳÐ໚¹Ç§¡ÅÁ (Closed
circular) 1 ǧ ÍÂً㹠Cytoplasm
à¡ÅÕÂǤً¢Í§àʌ¹ DNA ¨ÐÁÕ¡ÒúԴµÑÇã¹ÅѡɳÐ
"Supercolied" Relaxed circle
ÅѡɳСÒèѴµÑǢͧ Chromosome ໚¹ÃٻËҧ
·Õèä Á‹á¹‹¹Í¹ àÃÕÂ¡Ç‹Ò "Nucleoid"
Nucleoid = DNA (~60%) + RNA (mRNA) +
Protein (Nucleoid prot., Transcription factor)

4
 Eukaryote
Chromosomal DNA ÍÂً㹠Nucleus
ÍÂًã¹ÅѡɳТͧ DNA-protein complex ·Õè
àÃÕÂ¡Ç‹Ò "Chromatin"
Protein ·Õèà¡ÒÐÍÂً¡Ñº DNA ʋǹãË­‹ ¤×Í Histone
Histones ÁÕ 5 ª¹Ô´: H1, H2A, H2B, H3, H4
DNA ·Õ¾è ѹÃͺ Histone 8 ˹‹Ç 䴌ŠѡɳÐ໚¹
àÁç´ ·ÕèÁÕàʌ¹¼‹Ò¹Èٹ¡ ÅÒ§ ~10 nm àÃÕ¡NjÒ
"Nucleosome"

5
 RNA
RNA ໚¹ polynucleotide ÊÒÂà´ÕÂè Ç
ʋǹ»ÃСͺ¤ÅŒÒ¢ͧ DNA ¤×Í »ÃСͺ´ŒÇÂ
RiboNucleicAcid Phosphate group, Ribose áÅÐ Nitrogen bases
RNA µ‹Ò§¨Ò¡ DNA ¤×Í
¹éíÒµÒÅ໚¹ Ribose
Base Uracil (U) á·¹ Thymine (T)

Major type of RNAs mRNA

RNA ·Õºè ÃèآŒÍÁÙÅ "Message" ·Ò§¾Ñ¹¸Ø¡ÃÃÁ ¨Ò¡
Messenger RNA (mRNA) DNA à¾×èÍãˌ Ribosome áÅÐ tRNA ª‹Ç Â㹡ÒÃá»Å
ÃËÑÊ à»š¹ Protein
Transfer RNA (tRNA) "Message" ¨ÐºÃèØÍÂً㹡Å؋Á ¢Í§ Nucleotides 3 µÑÇ
Ribosomal RNA (rRNA) «Öè§àÃÕÂ¡Ç‹Ò “Codons”
ᵋŠРCodon ¨ÐÁÕ¤ÇÒÁ¨íÒà¾ÒСѺª¹Ô´¢Í§ Amino acid
ã¹ÊÒ Protein â´Â¨Ð໚¹ä»µÒÁ¡¯¢Í§ "Genetic
code"

6
 tRNA rRNA
tRNA ÁÕ “Anti-codon” «Ö§è ¡ç¤Í× ÅíҴѺ¡ÒÃàÃÕ§µÑǢͧ ໚¹Í§¤ç»ÃСͺ¢Í§ Ribosomes «Ö§è ÁÕº·ºÒ·ÊíÒ¤Ñ-ã¹
Nucleotides 3 µÑÇ ·ÕÊè ÒÁÒöࢌҤً¡Ñº "Codon" º¹àʌ¹ ¡Ãкǹ¡Òà Translation
mRNA
»ÅÒ´ŒÒ¹ 3' ¢Í§ tRNA ¨Ðà¡ÒСѺ Amino acid ·Õè
¨íÒà¾ÒÐ
´Ñ§¹Ñ¹é tRNA ¨Ö§à»š¹µÑÇàª×Íè ÁÃÐËNjҧÅíҴѺàºÊ·Õè໚¹ÃËÑʺ¹
àʌ¹ mRNA áÅÐÊÒ¢ͧ Amino acid ÊíÒËÃѺ
¡Ãкǹ¡Òà Translation à¾×èÍãˌ䴌 Protein

Replication Translation

DNA mRNA Protein

Transcription

7
DNA Replication

Replication
Semi-conservative = ¡ÒÃÊѧà¤ÃÒÐË• DNA ãËÁ‹
â´Â¡Ò÷Õè DNA à´ÔÁ«Öè§à»š¹à¡ÅÕÂǤًÍÂً¹Ñé¹ á¡ÊÒÂ
ÍÍ¡ÁÒ໚¹àʌ¹à´ÕèÂÇ¡‹Í¹ ¨Ò¡¹Ñé¹áµ‹ÅÐÊÒ·íÒ
˹ŒÒ·Õè໚¹áÁ‹áºº (Template) ã¹ ¡ÒÃÊѧà¤ÃÒÐË•
polydeoxyribonucleotide ÊÒÂãËÁ‹¢Öé¹ÁÒ ¼Å·Õè䴌
¤×Í DNA ·ÕèÊÌҧ¢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 (àʌ¹¤Ù‹) ¤ÅÒÂà¡ÅÕÂÇ໚¹ÊÒÂà´ÕèÂÇ ·Õè oriC
(Helicase, Single-stranded DNA binding
proteins (SSBs), DNA Gyrase)
ÊÌҧ RNA primer ÊÒÂÊÑé¹ æ (Primase)
µ‹ÍÊÒ polynucleotides ãˌÂÒÇ¢Öé¹ â´Â¡ÒÃàµÔÁ
dNTPs (5'3') ·Õè»ÅÒ¢ͧ RNA primer â´Â㪌
ÊÒÂà´ÔÁ໚¹áÁ‹áºº (DNA polymerase III)
DNA polIII ¨ÐÊѧà¤ÃÒÐË• DNA ã¹·ÔÈ·Ò§¨Ò¡ 5'3'
´Ñ§¹Ñé¹ DNA 2 àʌ¹·ÕèÊÌ Ò§¢Öé¹ãËÁ‹¨Ðµ‹Ò§¡Ñ¹

Mechanisms of Replicationtion in E. coli Enzymes participate in Replication

DNA ´ŒÒ¹·ÕèÊÒÁÒöÊѧà¤ÃÒÐˏ໚¹àʌ¹ÂÒǵ‹Íà¹×èͧ
¡Ñ¹ (Leading strand)
DNA ´ŒÒ¹·ÕèÊѧà¤ÃÒÐˏ䴌໚¹ DNA ÊÒÂÊÑé¹ æ
(Lagging strand, Okazaki fragment)
Lagging strand: àÁ×èÍ DNA polIII ÊÌҧ DNA ÊÒÂ
ãËÁ‹¢Öé¹ÁÒ¨¹ÁÒª¹¡Ñº RNA primer ·ÕèÍÂً¡‹Í¹Ë¹ŒÒ
DNA polIII ¨ÐËÅØ´ÍÍ¡ ¨Ò¡¹Ñé¹ DNA polI ¨ÐࢌÒÁÒ
‹Í RNA primer ·Ôé§áŌÇÊÃŒÒ §ÊÒ DNA ŧ᷹
DNA ligase ·íÒ˹ŒÒ ·Õèàª×èÍÁ DNA ÊÍ§Ê‹Ç¹à¢ŒÒ ´ŒÇ¡ѹ
Helicase & Single strand binding proteins
¤ÅÒÂà¡ÅÕ Ǥً¢Í§àʌ¹ DNA à¾×èÍàµÃÕ Áänj໚¹
áÁ‹áººã¹¡ÒÃÊѧà¤ÃÒÐˏ DNA àʌ¹ãËÁ‹
Primase Êѧà¤ÃÒÐË• Primer à¾×èÍ㪌 ໚¹Ê‹Ç¹àÃÔèÁ µŒ¹
¢Í§¡ÒÃÊѧà¤ÃÒÐË•ÊÒ DNA
DNA pol III µ‹Í¤ÇÒÁÂÒǢͧàʌ¹ DNA
DNA pol I ‹Í RNA primer ÊÃŒÒ § DNA ¢Öé¹á·¹
DNA ligase àª×èÍÁªÔé¹Ê‹Ç¹¢Í§ÊÒ DNA ࢌҴŒÇ¡ѹ

9
 RNA synthesis

Transcription
Ê‹Ç ¹·ÕèàËÁ×͹ DNA replication
ÁÕ¡ÒÃàµÔÁ dNTPs ã¹·ÔÈ·Ò§¨Ò¡ 5’ ä» 3’
Ê‹Ç ¹·Õ赋ҧ¨Ò¡ DNA replication
㪌ᤋºÒ§Ê‹Ç ¹¢Í§ DNA ໚¹áÁ‹áºº (template)
㪌 RNA polymerase 㹡ÒÃàµÔÁ dNTPs
àÁ×Íè àÊÃç¨ÊÔ¹é ¡Ãкǹ¡Òà ¨Ð䴌 single stranded RNA
Transcription
㪌 RNA polymerase 㹡ÒÃÊѧà¤ÃÒÐË• RNA ¨Ò¡
DNA
Template ¤×Í ÊÒ“Anti-sense”
µíÒá˹‹§àÃÔÁè µŒ¹¡Òà Transcription ·ÕèÍÂًº¹àʌ¹
Template ¤×Í “Promoter region”
¨Ø´ÊÔ¹é ÊØ´¢Í§¡Òà Transcription ¤×Í “Terminator
region”

10
 Mechanisms of Transcription
RNA pol ¨Ðà¢ŒÒ ¨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

11
 Translation
¡ÒÃá»Å¢ŒÍÁÙÅ·ÕèºÃèØÍÂًº¹ mRNA ä»à»š¹ÅíҴѺ¢Í§ amino
acid ·Õ¨è Òí à¾ÒÐ (Protein)
¡Ãкǹ¡Òà Polymerization ¢Í§ amino acids ä»
໚¹ Polypeptide sequence â´ÂÁÕâÁàÅ¡ØŢͧ
Ribosome áÅÐ tRNA ÁÒà¡ÒСѺ mRNA à¾×èÍ¡‹Íãˌà¡Ô´
¡ÒÃÊѧà¤ÃÒÐË•ÊÒ Polypeptide
Ribosome
The major participant in Translation
mRNA: ºÃÃ¨Ø Codon (3nt.), µíÒá˹‹§¨´¨íҢͧ
Ribosome, Start & Stop signals ÊíÒËÃѺ
¡ÒÃá»ÅÃËÑÊ໚¹ protein
tRNA: ºÃÃ¨Ø Anti-codon (ÁÕ¤ÇÒÁ¨íÒà¾ÒСѺ codon)
Ribosome: à˧»¯Ô¡ÔÃÔÂÒ¡ÒÃÊÌҧ¾Ñ¹¸Ð Peptide
ÃÐËNjҧ Amino acid ã¹ÊÒ Protein
Prokaryote Eukaryote
12
 tRNA Genetic code

Mechanisms of Translation Initiation

Initiation
Ribosome ࢌÒÁÒà¡ÒСѺ mRNA áÅзÕèµíÒá˹‹§
Start codon tRNA ¨Ð¹íÒ Amino acid µÑÇááࢌÒ
ÁÒà¾×èÍàÃÔèÁ¡Ãкǹ¡ÒÃÊÌҧÊÒ Polypeptide
Elongation
Ribosome à¤Å×è͹·Õäè »µÒÁÊÒ mRNA ¾ÃŒÍÁ¡Ñº¡ÒÃ
á»ÅÃËÑÊ Codon áÅйíÒ Aminoacly-tRNA ·ÕèÁÕ Anti
codon µÃ§¡Ñ¹ÁҨѺ à¾×èÍ¢ÂÒÂÊÒ Polypeptide
ÍÍ¡ä»
Termination
·ÕèµíÒá˹‹§ Stop codon ÊÒ Polypeptide ¨ÐËÂØ´
¡ÒÃÊÌҧ

13
Elongation Termination

14
 Genetic regulation

‘OPERON’ Lactocs ( lac) operon

¡ÅØÁ‹ ¢Í§ Genes ·Õè·íÒ˹ŒÒ·ÕèÃNj Á¡Ñ¹ã¹ Prokaryotes
â´ÂÁÕ»˜¨¨Ñ·Õè¨ÐÁҤǺ¤ØÁãˌ Genes àËŋҹÑé¹ÁÕ¡ ÒÃáÊ´§ÍÍ¡
à¾×èÍ·íÒ˹ŒÒ·Õè¨íÒà¾ÒÐÀÒÂã¹à«Åŏ
Operons ¨Ð·íÒ˹ŒÒ·Õèã ¹¡Òà ‘Induce’ ËÃ×Í ‘Repress’
¡ÒÃÊѧà¤ÃÒÐË• Protein â´ÂÁÒ¡ÁÑ¡¤Çº¤ØÁ·Õ¢è ¹Ñé ¢Í§¡ÒÃ
Transcription
¨Ð¾º¡Ò÷íÒ§Ò¹¤ÅŒÒÂ æ ¡Ñ¹¹Õãé ¹ Eukaryotes ᵋ¨ÐÁÕ¤ÇÒÁ
«Ñº«ŒÍ¹¡Ç‹Ò

15
Lactocs (lac) operon Lactocs (lac) operon

RNA Polymerase
RNA Polymerase

Mutation
Mutation ¡ÒÃà»ÅÕè¹á»Å§·Õèà¡Ô´¢Öé¹ã¹ÊÒþѹ¸Ø¡ÃÃÁ ໚¹¼Å
ãˌÁÕ¡ÒÃáÊ´§ÅѡɳзÕèà»ÅÕèÂ¹ä» áÅÐÊÒÁÒö¶‹ÒÂ
·Í´ÅѡɳйÑé¹ä»ÊًÅÙ¡ËÅҹ䴌

16
Causes of Mutation Types of Mutation

Spontaneous Base-pair insertion

à¡Ô´¢Ö¹é àͧ â´ÂÁÕ¡ÒÃà»ÅÕ¹á»Å§áººäÁ‹¨íÒà¾ÒÐà¨ÒШ§
Base-pair deletion
º¹àʌ¹ DNA
Induced Base-pair substitution
¶Ù¡¡Ãе،¹ â´ÂÊÀÒÇÐ ËÃ×ÍÊÒ÷Õè·íÒãˌà¡Ô´¡ÒáÅÒ¾ѹ¸Ø
‘mutagens’ ઋ¹ ÃѧÊÕ ÊÒÃà¤ÁÕ

Insersion causing Framshift Deletion causing Framshift

17
 Insertion or Deletion (No Framshift)

Substitution (Silent mutation) Substitution (Missence mutation)

18
 Substitution (Nonsence mutation)

Gene transfer

Types of Genetic exchanges

Transformation
Transduction
Conjugation

19
Transformation
¡ÒÃà¤Å×è͹ŒÒÂÊÒþѹ¸Ø¡ÃÃÁ (DNA) ¨Ò¡áº¤·ÕàÃÕÂà«ÅÅ•
˹֧è ä»ÂѧẤ·ÕàÃÕÂÍÕ¡à«Åŕ˹֧è
à«Åŏ·ÕèÃѺÊÒþѹ¸Ø¡ÃÃÁµŒÍ§ÁÕÊÀÒ¾·Ò§ÊÃÕÃÇÔ·ÂÒ·Õè àËÁÒÐÊÁ
àÃÕ¡à«ÅÅ•·ÃÕè ºÑ DNA ¹ÕéÇ ‹Ò "Competent"
à¡Ô´ã¹áº¤·ÕàÃÕª¹Ô´à´ÕÂǡѹ ᵋµ‹Ò§ÊÒ¾ѹ¸Ø ËÃ×Íà¡Ô´ã¹
Ấ·ÕàÃÕµ‹Ò§ª¹Ô´¡Ñ¹¡ç䴌
Ấ·ÕàÃÕÂ㹸ÃÃÁªÒµÔ·àÕè ¡Ô´ Transformation 䴌§‹Ò ઋ¹
Acinetobacter, Azotobacter , Bacillus ,
Streptococcus, Haemophilus

Transduction

¡ÒÃà¤Å×è͹ŒÒÂÊÒþѹ¸Ø¡ÃÃÁ (DNA) ¨Ò¡áº¤·ÕàÃÕÂà«ÅÅ•

˹֧è ä»ÂѧẤ·ÕàÃÕÂÍÕ¡à«Åŕ˹֧è â´ÂÍÒÈÑ Bacterial
virus (Bacteriophage)
à¡Ô´ã¹áº¤·ÕàÃÕª¹Ô´à´ÕÂǡѹ

20
Conjugation
¡ÒÃࢌҤً¡ ѹ¢Í§áº¤·ÕàÃÕ áÅÐÁÕ¡ Òö‹Â·Í´ÊÒþѹ¸Ø¡ÃÃÁ
(DNA) ãˌᡋ¡ ѹ â´ÂẤ·ÕàÃÕÂà«Åŏ˹Ö觨Ð໚¹¼ÙŒãˌ
(Donor) áÅÐÍÕ¡à«Åŏ˹Öè§à»š¹¼ÙŒÃѺ (Recipient)
à«ÅÅ•¼ÙŒã ˌ¨ÐÁÕ F plasmid (F+) áÅÐÊÒÁÒöÊÌҧ
"Sex pilus" à¾×èÍ㪌㹡ÒèѺ¤ÙዠÅÐʋ§ DNA ࢌÒä»ã¹à«Åŏ
¼ÙÌ Ѻ (F-) J. Lederberg & E. Tatum (1958)
à¡Ô´ã¹áº¤·ÕàÃÕ¡Å؋Á·ÕèÊÒÁÒöࢌҤً¡Ñ¹ä´Œ

21
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