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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