Fig. 12-00. Fig. 12-01 Fig. 12-02 Fig. 12-03 Fig. 12-04.

Fig. 12-00

Fig. 12-01

Fig. 12-02

Fig. 12-03

Fig. 12-04

Fig. 12-05

Fig. 12-06

Fig. 12-07

Plasmids

Bacterialchromosome

Remnant ofbacterium

Co

lori

zed

TE

M

Fig. 12-08-1

Plasmid

Bacterial cell

Isolateplasmids.

Fig. 12-08-2

Plasmid

Bacterial cell

Isolateplasmids.

DNA

IsolateDNA.

Cell containingthe gene of interest

Fig. 12-08-3

Plasmid

Bacterial cell

Isolateplasmids.

DNA

IsolateDNA.

DNA fragmentsfrom cell

Cut both DNAswith sameenzyme.

Gene ofinterest

Othergenes


Fig. 12-08-4

Plasmid

Bacterial cell

Isolateplasmids.

Gene of interest

Recombinant DNA plasmids

DNA

IsolateDNA.



Gene ofinterest

Othergenes

Mix the DNAs andjoin them together.


Fig. 12-08-5

Plasmid

Bacterial cell

Isolateplasmids.

Recombinant bacteria

Gene of interest


Bacteria take up recombinant plasmids.

DNA

IsolateDNA.



Gene ofinterest

Othergenes



Fig. 12-08-6

Plasmid

Bacterial cell

Isolateplasmids.

Clone the bacteria.

Recombinant bacteriaBacterial clone

Gene of interest



DNA

IsolateDNA.



Gene ofinterest

Othergenes



Fig. 12-08-7

Plasmid

Bacterial cell

Isolateplasmids.

Find the clone withthe gene of interest.

Clone the bacteria.


Gene of interest



DNA

IsolateDNA.



Gene ofinterest

Othergenes



Fig. 12-08-8

Plasmid

Bacterial cell

Isolateplasmids.

Some usesof genes

Gene for pestresistance

Gene fortoxic-cleanupbacteria

Genes may beinserted intoother organisms.

Find the clone withthe gene of interest.

The gene and proteinof interest are isolatedfrom the bacteria.

Clone the bacteria.


Gene of interest



Harvestedproteins may beused directly.

Some usesof proteins

Protein for“stone-washing”jeans

DNA


Protein fordissolvingclots

IsolateDNA.



Gene ofinterest

Othergenes


Fig. 12-09-1 Recognition sequencefor a restriction enzyme

Restrictionenzyme

Sticky

end

Stickyend

DNA

A restriction enzyme cutsthe DNA into fragments.


Restrictionenzyme

Sticky

end

Stickyend

DNA

A DNA fragment is addedfrom another source.



Restrictionenzyme

Sticky

end

Stickyend

DNA



Fragments stick together bybase pairing.


Restrictionenzyme

Sticky

end

Stickyend

DNA

DNAligase

Recombinant DNA molecule



Fragments stick together bybase pairing.

DNA ligase joins thefragments into strands.

Fig. 12-10

Radioactive probe(single-stranded DNA)

Single-stranded DNA

Mix with single-stranded DNA fromvarious bacterial clones

Base pairing indicates thegene of interest

Fig. 12-11-1Cell nucleus

DNA ofeukaryoticgene

Test tube

Transcription

Exon Intron Exon ExonIntron



RNAtranscript

mRNA

Test tube

Transcription

Introns removed andexons spliced together




RNAtranscript

mRNA

Test tube

Reversetranscriptase

Transcription


Isolation of mRNA fromcell and addition ofreverse transcriptase




RNAtranscript

mRNA

Test tube


cDNA strandbeing synthesized

Transcription



Synthesis of cDNAstrand




RNAtranscript

mRNA

Test tube

cDNA of genewithout introns


cDNA strandbeing synthesized

Transcription



Synthesis of cDNAstrand

Synthesis of second DNAstrand by DNA polymerase


Fig. 12-12

Fig. 12-13-1

DNA isolated

Crime scene Suspect 1 Suspect 2

Fig. 12-13-2

DNA isolated

DNA amplified


Fig. 12-13-3

DNA isolated

DNA amplified

DNA compared


Fig. 12-14

Fig. 12-14a

Fig. 12-14b

Fig. 12-15

InitialDNAsegment

Number of DNA molecules

1 2 4 8

Fig. 12-16

Crime scene DNA

Suspect’s DNA

Same number ofshort tandem repeats

Different numbers ofshort tandem repeats

STR site 1 STR site 2

AGAT

AGAT GATA

GATA

Fig. 12-17-1

Mixture of DNAfragments ofdifferent sizes

Powersource

Gel

Fig. 12-17-2


Powersource

Gel

Fig. 12-17-3


Powersource

Gel

Completed gel

Band oflongest(slowest)fragments

Band ofshortest(fastest)fragments

Fig. 12-18

Amplifiedcrime sceneDNA

Amplifiedsuspect’sDNA

Longerfragments

Shorterfragments

Fig. 12-19

Crime sceneDNA

Suspect’sDNA

Fragment w

Fragment x

Fragment y

Longerfragments

Shorterfragments

Fragment z

Fragment y

Crime sceneDNA

Suspect’sDNA

Cut

Cut Cut

Restriction enzymes added

x

wy y

z

Fig. 12-20

Fig. 12-21

Anthraxspore

Envelopecontaininganthrax spores

Fig. 12-22-1

Chromosome

Fig. 12-22-2

Chromosome

Chop up withrestriction enzyme

DNA fragments

Fig. 12-22-3

Chromosome


Sequencefragments

DNA fragments

Fig. 12-22-4

Chromosome


Sequencefragments

DNA fragments

Alignfragments

Fig. 12-22-5

Chromosome


Sequencefragments

DNA fragments

Alignfragments

Reassemblefull sequence

Fig. 12-22a

Fig. 12-23

Fig. 12-24-1

Normal humangene isolatedand cloned

Healthy person

Fig. 12-24-2


Normal humangene insertedinto virus

Healthy person

Harmlessvirus (vector)

Virus containingnormal human gene

Fig. 12-24-3


Normal humangene insertedinto virus

Virus injectedinto patient withabnormal gene

Healthy person

Harmlessvirus (vector)

Virus containingnormal human gene

Bonemarrow

Bone of personwith disease

Fig. 12-25

Fig. 12-26

Fig. 12-27

Fig. 12-28

Fig. 12-T01

Fig. 12-UN01

DNA isolated from twosources and cut by samerestriction enzyme

Gene of interest(could be obtained froma library or synthesized)

RecombinantDNA

Plasmid(vector)

Transgenic organisms

Useful products

Fig. 12-UN02


DNA

Polymerase chainreaction (PCR)amplifies STRsites

LongerDNAfragments

ShorterDNAfragments

DNA fragments compared by gel electrophoresis

Gel

Fig. 12-UN03

Normalhuman gene

Virus

Bonemarrow

Normal human gene is transcribedand translated in patient, potentiallycuring genetic disease permanently

Fig. 12-00. Fig. 12-01 Fig. 12-02 Fig. 12-03 Fig. 12-04.

Documents

Transcript of Fig. 12-00. Fig. 12-01 Fig. 12-02 Fig. 12-03 Fig. 12-04.