presentasi rekayasa genetika

8/17/2019 Presentasi Rekayasa Genetika

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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 .



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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



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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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