Slide Dna Recom

8/17/2019 Slide Dna Recom

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PREPARED BY :MARWIN KUMARPREPARED BY :MARWIN KUMAR

:NEOH KAI CHYUEN:NEOH KAI CHYUEN

:ELEANORA WONG:ELEANORA WONG

:SYAFIQULLAH KHAN:SYAFIQULLAH KHAN :TAN JING EE:TAN JING EE

:SITI NUR ZULAIQHA:SITI NUR ZULAIQHA

LECTURER :MADAM NOR HAYATILECTURER :MADAM NOR HAYATI

SUBJECT :LIFE SCIENCESSUBJECT :LIFE SCIENCES


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RDNA/ RDNA tech Restriction enzymes/endonucleases

Plasmid DNA ligase Vectors

Bacteriophages DNA library


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DNA that has been created artificially (not

natural) DNA from t!o or more sources is

incorporated into a single recombinant molecule


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Procedures by !hich DNA from differentspecies can be isolated" cut and splicedtogether

Ne! #recombinant # molecules are thenmultiplied in $uantity in populations of rapidlydi%iding cells (eg bacteria" yeast)

&ses methods deri%ed from biochemistry of

nucleic acids coupled !ith genetic techni$uesoriginally used for the study of bacteria and%iruses


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' A gene is located on a chromosome map A DNA library is constructed plasmid are

obtained" clea%age occurs and RDNA prod * +he gene of interest is isolated (cloned) from

the library %ia plasmid DNA isolation" restrictiondigestion and electrophoresis

, -ultiple copies of gene interest are producedfor study


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Restriction enzymes .cuts DNA

at specific sites (restriction fragments)

DNA ligase .pastes the DNAfragments together

+he cut segments are inserted into other DNA+he cut segments are inserted into other DNA

molecules that ser%es asmolecules that ser%es as %ectors(P0A1-2D)


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DNA cutting enzymes Restrictionendonucleases

cuts DNA at a specific site defined by ase$uence of bases in the DNA (recogsite)

forming .stic3y ends (ss) Palindromic sites 4g Bam52 cuts 67 88A+99 *7

*7 99+A886: 4g 5ae222 cuts 678899*7*7998867

se%eral hundred endonucleases ha%e beene;tracted from bacteria and many are used inrecombinant DNA research eg 4coR'"5ind 222"5ae222" +a$A'" 1au*A


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BACTERIAL ORIGIN=ENZYME WHICH CLEAVE FOREIGN DNA AS

IT(ECORI;ESCHERICHIA COLI) PROTECT BACTERIA FROM

BACTERIOPHAGE INFECTION


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9arrier DNA mol < transfers the RDNA into thehost cell

=ithin host cells" %ectors can replicate producingmany DNA segments < identical copies

(90>N41) 5ost cells pass the recombinant DNA mol on their

progeny < population of cells 9loned DNA segments reco%ered from host cells

for purification and analysis


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-olecules of DNA that are found in bacteria

Act as a system to transfer genetic material to

other bacteria" allo!ing those to e;press thetransmitted genes

small (a fe! thousand base pairs)

usually carry only one or a fe! genes

circular

ha%e a single origin of replication


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•Schematicrepresent typical

plasmic vector

which can e !se

"or w# screenin$

test.

•Typical w# screenin$ proce%!re


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Because each cloned DNA segement is relati%elysmall" many separate clones must be constructed

A set of cloned DNA segments deri%ed from asingle indi%idual represents a library

9loned libraries could be an entire genome" asingle chromosome" or a set og genes compiledtogether in a single cell type


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Applications of recombinant DNAApplications of recombinant DNA

technologytechnology

&sed !idely in research and hospital laboratories&sed !idely in research and hospital laboratories

Broad applications ? medicine" agriculture"Broad applications ? medicine" agriculture"


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-edicine production of industrial and commercial compounds 2nsulin Diabetes drugs (angiostation and endostatin) " @actor V222 5aemophilia A @actor 2 5aemophilia B

4P> Anaemia 2nterleu3ins and interferons +issue plasminogen acti%ator dissol%e blood clots 5ormones < 85" parathyroid >;ytocin Adenosine deaminase se% 9om 2mm (192D)

,


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Diagnostic 3itsDiagnostic 3its 5ep" A2D1 AntibioticsAntibiotics

genetic testing" -apping the chromosomallocation of genetic disorders R@0P" DNA

fingerprinting the 58 proect 8ene therapy manipulation of DNA to t;

diseases by altering indi%iduals genes (9@"PC&" D-D) 4+529A0 211&41

@orensic applications ? All indi%iduals aregenetically uni$ue < a distinct #geneticfingerprint." all types of specs" old and ne!

Animals < models of genetic diseases 4g 84 mice


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Vaccines %Vaccines %are created by transferring thegenes that determine a pathogen7s surfaceconfiguration to a microorganism

=hen the 8- microorganism is used in a%accine" its surface stimulates the production ofantibodies

+hese antibodies protects an indi%idual against

the pathogen 1uccessful %accines ha%e been produced for

influenza" cold sores" and hepatitis B(5BsAg)


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Animals impro%ed $uality/$uantityof meat products

Plants 8ene altered plants impro%ed crops $uantity" $ualityand pest resistance 4g golden rice 8- to contain beta carotene %it Adeficiency

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