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Ff bacteriophage
E.coli
PS
ssDNA
Gene
TolA expression
F pilus
dsDNA (RF)
pIII + pVI pVIII pI/pXI + pIV

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Replication Virion Assembly/export
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Displayed Displayed
sequence sequence
TAG
Sig. Gene III Sig. Gene III
Phage Phagemid
vector vector
pBR322 ori
Antibiotic Ff ori Antibiotic Ff ori

resistance resistance
gene–optional gene
E. coli E. coli
(Defective
Ff ori)
Add helper phage
Polyvalent Monovalent
phage phage
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Construct vector and

confirm functional display General principles: Ch. 4
General principles: Ch. 4

Oligo-directed mutagenesis: Ch. 2
In vitro recombination: Ch. 3
Oligo cassette: Ch. 6
Randomize targeted regions Recursive PCR: Ch. 8
Oligo-splinted assembly: Ch. 9
Error-prone PCR: Ch. 11
PCR with randomized primers: Ch. 14
Obtain or Premade peptide libraries: Ch. 1 sect. 7

Gene fragment libraries: Ch. 11
create random
cDNA libraries: Ch. 12
gene library Antibody V-region libraries: Ch. 13
General principles: Ch. 4

In vitro binding: Chs 4, 6, 9, 11, 12, 13
Select Enzymatic selection: Chs. 7, 8
In vivo binding: Ch. 10
Sequencing: Ch. 4 sect. 7

Analyse PCR fingerprinting: Ch. 13
selected Binding, by phage ELISA: Ch. 5
clones (also see Chs. 6, 9, 11, 12, 13)
Test soluble proteins: Chs. 4, 6, 13, 14
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N54%.-#%D' TeR8 77 FKKK XReZ

J+)4"4-&%2' ?\4#"2*0'#"*' XJ?,Z 8 & 8e@e FKKK XRfZ

6"! =023. (' 1023.?,) E.+&('F

Antibiotic Ff ori Antibiotic Ff ori

6"5 =(4>E24.%& (' ?(%(E24.%& ),:142>F

6"6 G.41.' 1023.

6"8 9.%.'24 1'(&(+(4: /(' 1023. 1'.12'2&,(% 2%)

L =&*&'#2 $'/*./$2&4 +7 # $"#0& ,/4$2#3 $'+F&.%

8"# I2J,%3 2 4,@'2'>

Construct vector and

General principles: Ch. 4

Obtain or Premade peptide libraries: Ch. 1 sect. 7

General principles: Ch. 4

Sequencing: Ch. 4 sect. 7

8"5 K%24>:,: (/ +4(%.:

<#12& : H]"DF+'* %0 .%2S&4&%2* )*'S 0%# '+)4&%2 %0 4"#I'4\T%)2S F-"I'

O2-%/+* 1-77&' K&7&'&*.&

8:b7:: D! AN+ XRe6 f9Z

ebfe D! E,, X>&4- #'S)."T+' +&2/'#Z Xee6 fP6 feZ

7:: D! 4#&'4-5+"D&2' XfRZ

e: D! *%S&)D .&4#"4'6 FA 8bR XfQZ

!"! -&'+&'. (/ &0. 1023. 12'&,+4.

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=&& >'+%&/ ?/@& A##B 9-.%/+ C+.#%/+* D4&, 7+' ,/4$2#3E

KK KK P:O ('F+&."4&%2 X2&./"'Z N54%F+"D

_KK _KK 99 _&#&%2 .%DF%2'24 _&#&%2 4&F X4"#4Z ;' X=\4'#DZ

Ka Ka 98 _&#&%2 .%DF%2'24 _&#&%2 4&F X4"#4Z ;' X=\4'#DZ

d%(-3&+"(- "-:(%:# +2 #155##": .$%&50-+ () $c ,"0.# +2&+ 5(:. +2

<#12& 5 H]"DF+'* %0 F#%4'&2* S&F+"5'S %2 W+"D'24%) F-"I'"

!)4"#S 4#5F&2 &2-&T&4%# X!,K\8Z f FKKK XPQZ

?)T4&+&&2 8Q FKKK6 F_KKK Xee6 eRZ

<+/"+&2' F-%F-"4"' 8 & R: FKKK XR7Z

J+)4"4-&%2' ?\4#"20'#"' XJ?,Z 8 & 8e@e FKKK XRfZ

L =&&'#2 $'/./$2&4 +7 # $"#0& ,/4$2#3 $'+F&.%

G#-7#.%-'&' <'#,&#8& V&.%+' K&#0&*%4 #T#/2#12& W&14/%&