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GENETIC ENGINEERING
Presented by:
1. Wahyu Maulana Endris 15725251010
2. Royan Mahmud Musthofa 15725251038
PROGRAM STUDI PENDIDIKAN BIOLOGIFAKULTAS PASCASARJANA
UNIVERSITAS NEGERI YOGYAKARTA2016
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Contents
1 !ISTORICAL OF GENETIC ENGINERING
2 Re"o#$%n&nt 'n& te"(no)o*+
, B&se te"(no)o*+ o- 'n& .e"o#$%n&nt
/ E%#ent .e"o#$%n&nt 'n& te"(no)o*+
3 Ce)))&. en4+#e
6 N&t.&) 5e"to.
Bene7t o- te"(no)o*+ .e"o#$%n&nt
8 Te"(n%es o- Genet%" En*%nee.%n*
9 Stes o- Genet%" En*%nee.%n*
10A)%"&t%on o- Genet%" En*%nee.%n* %n
D&%)+ L%-e
11B%oet(%"s
Wah u maulana endris 157252510
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1 !%sto.%"&)o- *enet%"
en*%ne.%n*
1!31Phenomena re"ombinationand #hysi"al e$iden"e %&'
(as found.
1!)8Endonu"lease (as dis"o$er or
en*yme restri"tion done byste(art linn and (erner arber.
1!73 +he ,rst e-#eriment inre"ombinant dna "lonindone by herbert boyerand stanley "ohen .
Wah u maulana endris 157252510
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1 !%sto.%"&)o- *enet%"
en*%ne.%n*
1!7!' method of#rodu"in insulin
usin eneti"enineerin (asfound .
1!80/irst mi"e eneti"ally
enineered (as de$elo#ed.
1!!1ene thera#y ,rst triedand tested in humans .
1!! +anential tomatoes released in
the maret for the ,rst time
1!!7%olly the ,rst animal"lone
Wah u maulana endris 157252510
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Re"o#$%n&ntDNA
Te"(no)o*+
• ' lon time ao our forefathers a(are ofdi$ersity li$in thins.
• %i$ersity this li$in thin has allo(ed humans to"hoose (hi"h ty#e of li$in thins he dire"ted .
• +e"hni4ue re"ombinant dna is a te"hni4ue ineneti"ally enineered to #rodu"e ne(
#ro#erties by means of Re"ombinin a #arti"ularene of the dna (ith the enome .
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Re"o#$%n&ntDNA
Te"(no)o*+
• +e"hnoloy no(n as re"ombinant dna
te"hnoloy or (ith the term more#o#ular is eneti" enineerin thisin$ol$in eort main "o#ies of a#arti"ular ene in a "ell that is not its
natural "ell buses often also (as said tobe "lonin ene.
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, B&sete"(no)o*+ o-
.e"o#$%n&nt 'n&
+ransfer dna or
dis#la"ement dna ordis#la"ement dna intoba"teria "an in three(ays namely "on6uationtransformation and
transdu"tion.
Con:*&t%o
T.&ns-o.#&t%o
T.&ns'"t%o
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Con:*&t%on
Resistant a"terium ensiti$e a"teriumResistant a"terium
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T.&ns-o.#&t%on
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T.&ns'"t%on
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/ Sets o- e%#ent -o. 'n&.e"o#$%n&nt te"(no)o*+
Rest.%"t%on En4+#e
• +rans#oson
• %na9rna dete"tor
• enom ma#
L%*&se En4+#e
E;+
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3 Ce)))&.en4+#e
• Rest.%"t%onEn'on")e&se
• DNA;RNAo)+#e.&se
• Dn& )%*&se
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3 Ce)))&.en4+#e
• .e5e.set.&ns".%t&se
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6 N&t.&)Ve"to.
'n instrument that"arries dna into hisne( stem "ells.
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Bene7t o-te"(no)o*+.e"o#$%n&nt
'##li"ation te"hni4ue
re"ombinant dna inbiote"hnoloy ofthem are $a""ine#rodu"tion insulin antibodies and forth.
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Te"(n%e o- DNA Re"o#$%n&nt
• %&' Re"ombinant i$e us ability to tae the s#e"i," enesfrom oranism "ombine those ene to other oranisminsert to the host "ell until the ability of "ell to e-#ress thoseind of "ombinin ene.
• +here are se$eral te"hni4ue of %&' re"ombinant
%&';ombini
n
%&';uttin
%&'>solation
%&'>nsertio
n
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DNA Iso)&t%on• +ain the %&' form the taret "ell Aenomi" %&' and
#lasmid %&'B
• ;rushin the "ell membrane or "ell (all by me"hani"al
or en*ymati"ally (ays
• ;rushin the nu"leus membrane by addin the +ritonCD100 or odium %ode"yl ul#hate A%B
• e#aratin the %&' from other "ell oranelles by
sentrifuation• Purifyin the %&' by addin the ammonium a"etate or
al"ohol
1
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D%&*. o- DNA Iso)&t%on
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DNA Ctt%n*• ;uttin the %&' by restri"tion endonu"lease
en*yme
•
Endonu"lease en*yme able to re"oni*inbindin and "uttin the restri"tion site.
• /or e-am#le E"oR> en*yme (ill "ut the %&' (hi"hs#e"i,"ally has the base se4uen"e of ''++;
• +he "uttin #ro"ess (ill result the asimetris #ointAstickey end) mean(hile the en*yme (hi"h "utthe %&' in blunt (blunt end)
2
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D%&*. o- DNA Ctt%n*
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St%"
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DNA Co#$%n%n*• +he im#ortant
"ondition of %&'"ombinin is the
enomi" %&' must be"om#atible (ith the$e"tor A#lasmidB %&'
• +hree (ays to"ombine the enomi"
%&' (ith the #lasmid%&'
3
FGsin the
%&' liaseen*ymefrom
ba"teria
HGsin the %&'
liase from E. Coli (hi"h hasinfe"ted by
ba"terio#hae +
Ii$in
deosinuleotidil
transferaseen*yme
to synthesi*esinle
strand ofhomo#olimeri"
3J
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D%&*. o- DNA Co#$%n%n*
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DNA Inse.t%on• %&' insertion also no(n as transformation found by M.
Mandel dan '. @ia A1!70B
• +he %&' re"ombinant and ba"teria Ahost "ellB mi- in the
solution of ;a;l2 in the tem#erature 0o K 5o;
• ;a;l2 solution (ill mae the ba"teria "ell to be s(ollen mae
the "ell (all lose the #eri#lasmi" #rotein and ,nally lea
• +he re"ombinant %&' (ill bind (ith ;a2L formin the %&'se"om#le- resistant
• +hose "om#le- (ill ,nally enter the ba"teria by heatCsho"treatment
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D%&*. o- DNA Inse.t%on
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T.&ns-o.#&nt Se)e"t%on• +he sele"tion of the "ell (hi"h "ontain the $e"tor (hi"h "arry
the %&' re"ombinant "an be dete"ted usin the tra"eCframent A probe)
•
+he main of probe usin Polymerase ;hain Rea"tion AP;RBby in $itro
• +he te"hni4ue to dete"t the host "ell "ontain desired eneusin "olony hybridi*ation.
• +he %&' of "ell sun in the tra"e solution so the tra"eC
frament A probeB (ill insert to the %&'• &e-t the #osition of %&' (hi"h inserted by the probe (ill
"om#are to the "olony #osition of initial "ulture Amaster plate)
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e"ause of not all "ell Aba"teriaB "ontain there"ombinant %&' so (e must sele"t the "ellthat "arry the re"ombinant %&'. +here are 3#ossibilities that "an ha##en after
transformation:
+he host "ell didnJt inserted by anyother ene
N +he host "ell reliation
O +he host "ell inserted by re"ombinant$e"tor (ith or (ithout the e-#e"tedene
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STEPS o- DNA Re"o#$%n&t%on AsummaryB
>solation of %&'
N ;uttin %&'
O ;ombinin %&'
'naly*e %&'
Q+ransformant and re"ombinantsele"tion
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ST
EPSo-DNARe"o#$%&t%onAsummaryB
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A)%"&t%on o- Genet%" En*%nee.%n* %n D&%)+ L%-e
• +he de$elo#ment of eneti" enineerin is$ery ad$an"ed throuh the in$ention of
modifyin eneti"• +he resear"her has #rodu"ed mu"h more
#rodu"t and transeni" oranism byeneti" enineerin no(n as Genetically
Modifed Organism AM
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M&n+ Se"to.s !&5e A=e"te' $+ Genet%" En*%nee.%n*
1./arms
+ranseni" #lant that• +olerant to "hemi"al
substan"es• Resistant to diseases• @as the s#e"i,"
"hara"teristi"• 'ble to bind &2 from
the air autonomously•
@a$e a oodada#tation
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2. @orti"ulture and/orestry
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3. Pharma"y and >ndustry
M&nsulinB and #lasmid• Gse =iase to "o$alently bind frament and #lasmid
Inse.t P)&s#%' Into B&"te.%&3
• Modi,ed #lasmid mi- (ith the ba"teria "ulture• a"teria (ill tae the #lasmid throuh transformation
C)on%n* o- Gene &n' !ost Ce))• +he ba"teria as the result of transformation are #la"ed in the solid
medium (hi"h "ontain am#i"illin and suar ("-GalB
I'ent%-+ t(e C)one5
• Gsin the tra"eCframent A probe) nucleid acid probe
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D%&*.&#o-
M&
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. En$ironment
ome transeni" ba"teria (ill hel# bioremediation of #olluted "oast and ri$er
5 =a(s and
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5. =a(s and/orensi"
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B%oet(%"s
• >n the de$elo#ment of "ien"e theresear"her and te"hno"rat (hom #lune
into a"ti$ity of mole"ular bioloy andeneti" enineerin must i$in attentionto the rules and ethi"s.
• ioethi"s i$in the solution to sol$e the
"ontro$ersy in the e-#erimentde$elo#ment and the usae of humanand natural resour"es
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A N K Y
O U
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