Investigatory Project Biology

  • Upload
    tushar

  • View
    1.024

  • Download
    27

Embed Size (px)

Citation preview

  • 7/25/2019 Investigatory Project Biology

    1/45

    INVESTIGATORY PROJECT

    BIOLOGY

    NAME: TUSHAR GAUTAM

    CLASS: 12THA

    ROLL NO.: 13

    SCHOOL: R.P.V.V. SEC-10 DWARKA NEW DELHI-11007

  • 7/25/2019 Investigatory Project Biology

    2/45

    DNA Fingerprinting

    Unless they are identical twins, individuals haveunique DNA

    DNA !"#$%&'&"#("#$

    The name used for the unamiguous identifying techniquethat ta!es advantage of differences in DNA sequence

    The process of DNA fingerprinting egins y isolatingDNA from

    lood, semen, vaginal fluids, hair roots, s!in, s!eletalremains, or elsewhere

    2

  • 7/25/2019 Investigatory Project Biology

    3/45

    "olymerase #hain $eaction %"#$&

    'f there is only a small amount of DNA availale for DNA

    Fingerprinting

    augment the amount of DNA using a

    technique called PCR

    PCR is doing DNA replication in a test

    tube

  • 7/25/2019 Investigatory Project Biology

    4/45

    Template

    and coolto anneal

    35

    Primer

    PrimerTemplate

    Like ALL DNApolymerases 53

    Taq polymerase canonly add to the 3 end

    of an existing

    nucleotide

    A DNA primer that is

    complementary to the

    template is used to

    supply that 3 end

    4

  • 7/25/2019 Investigatory Project Biology

    5/45

    DNA Fingerprinting

    After we isolate the DNA and amplify it with "#$ Treat the DNA with &%)(&"*("+# %#,%)

    cut DNA at specific sequences

    (veryone)s DNA is different, so everyone)s DNA will cut at different sites

    This results in different si*ed fragments

    The different si*ed fragments are called &%)(&"*("+# !&/$%#(%#$( '++&'")), or RLP)

    +e can oserve the different si*ed fragments in an eperiment

    that separates DNA ased on fragment si*e called -el(lectrophoresis

  • 7/25/2019 Investigatory Project Biology

    6/45

    $F." Analysis

    (veryone has geneticsequences called/&"/4% #54%&

    (/#6% &%'%/(), orVNTR)

    (veryone has differentamounts of /NT$s

    The /NT$s ma!e the

    different si*ed $F."s

    6

  • 7/25/2019 Investigatory Project Biology

    7/45

    -el (lectrophoresis

    Fragments of DNA from restriction en*yme cleavageare separated from each other when they migratethrough a support called an /$/&+)% $%

    't is similar to the yummy food 0ell12 gelatin

    't is actually made out of some of the same ingredients

    The si*e1ased separation of 3olecules of DNAseparate ased on si*e when an electric current isapplied to an agarose gel

    This is $% %%*(&+'+&%)")

    7

  • 7/25/2019 Investigatory Project Biology

    8/45

    -el (lectrophoresis

    8

  • 7/25/2019 Investigatory Project Biology

    9/45

    -el (lectrophoresis

    The separated DNA fragments are then drawn out of

    the gel using a nylon memrane

    The nylon memrane is treated with chemicals that

    rea! the hydrogen onds in DNA and separate thestrands

    The single stranded DNA is cross lin!ed to the nylonmemrane

    4y heat or U/ light

    9

  • 7/25/2019 Investigatory Project Biology

    10/45

    -el (lectrophoresis

    'ncuate the nylon memrane with a radioactive '&+4%of single stranded DNA complementary to the

    /NT$s

    10

  • 7/25/2019 Investigatory Project Biology

    11/45

    -el (lectrophoresis

    -el (lectrophoresis

    11

  • 7/25/2019 Investigatory Project Biology

    12/45

    The radioactive proe shows up on photographicfilm

    4ecause as it decays it gives off light

    The light leaves a dar! spot on the film

    Different individuals have different patterns ofands these ma!e up the fingerprint

    -el (lectrophoresis

    12

  • 7/25/2019 Investigatory Project Biology

    13/45

    This rotocol is kno!n as "outhern #lotting5outhern 4lotting

    13

  • 7/25/2019 Investigatory Project Biology

    14/45

    DNA Fingerprinting

    14

  • 7/25/2019 Investigatory Project Biology

    15/45

    DNA fingerprints can e used to determine whichone fragments elong to which individual

    DNA Fingerprinting

    15

  • 7/25/2019 Investigatory Project Biology

    16/45

    DNA fingerprints of children should e similar to the thoseof parents

    DNA fingerprinting can show which individuals are theparents of specific children

    16

  • 7/25/2019 Investigatory Project Biology

    17/45

    Northern 4lot Analysis

    Northern lotting analy*es $NA much the sameway that 5outhern lotting does DNA6

    $NA is etracted from the cell, undergoes gelelectrophoresis, and is ound to a filter7

    8yridi*ation etween ound cellular $NA and a laeledproe occurs7 The si*es of the $NA fragments detectedy the proe can e determined

    Northern 4lot Analysis

    Northern lot analysis is used for determining6

    17

  • 7/25/2019 Investigatory Project Biology

    18/45

    The si*e%s& of m$NA encoded y a gene7 Northern lotshave shown that different m$NA species arise from the

    same region of DNA, suggesting differential use ofpromoters and terminators, and9or alternative m$NAprocessing7

    +hether a specific m$NA is present in a cell type, and ifso, at what levels7 -ene activity is measured in this way,

    and $NA sampling is widely used to study development,tissue speciali*ation, or the response of cells to variousphysiological stimuli7

    "roducing $ecominant "roteins

    18

  • 7/25/2019 Investigatory Project Biology

    19/45

    The first step in the production of rBGHprotein %or insulin&is

    to transfer the BGHgene %or human insulin gene& from the nucleus ofa cow cell %or human cell& into a acterial cell

    8ow do we do that::::

    ; steps are involved in turning a cow BGHgene into arecominant 4-8 %r4-8& gene in a acterial cell

    rBGHgene means that this product is genetically engineered

    with the rindicating recominant

    "roducing $ecominant "roteins

    < steps are involved in turning a cow BGHgene into arecominant 4-8 %r4-8& gene in a acterial cell

    19

  • 7/25/2019 Investigatory Project Biology

    20/45

    1. 3a!e lots of copies of the cow 4-8 gene in the la in atest tue

    2. #ut cow 4-8 gene with restriction en*ymes

    3. 'nsert this cow 4-8 gene into acterial DNA = r4-8

    4. 'n>ect the acterial DNA containing the r4-8 into acteria

    5. -row up lots of these genetically engineered acteria andpurify the r4-8 cow protein they are ma!ing

    C+#"#$ / G%#% U)"#$ B/*(%&"/

    5tep ?7 3a!e lots of copies of the 4-8 -ene

    20

  • 7/25/2019 Investigatory Project Biology

    21/45

    Use "#$ to amplify only the cow 4-8 gene fromthe cow chromosomes

    $ememer, "#$ is >ust replicating DNA in thelaoratory in a test tue

    (nd up with lots and lots of copies of the cow 4-8gene DNA in a test tue

    #loning a -ene Using 4acteria

    5tep @7 "repare the cow 4-8 -ene for inserting intoacterial DNA

    21

  • 7/25/2019 Investigatory Project Biology

    22/45

    The cow 4-8 gene ends are sliced using&%)(&"*("+# %#,%)

    $estriction en*ymes cut DNA only at specificsequences that leave the doule1stranded DNA>agged or stic!yB on the ends

    $estriction en*ymes cut the DNA in a staggered

    22

  • 7/25/2019 Investigatory Project Biology

    23/45

    pattern, leaving sticky ends

    Restrictin en!"#e c$t

    23

  • 7/25/2019 Investigatory Project Biology

    24/45

    Fig. 16.2Examples of how

    differentrestriction

    enzymes cleaveDNA

    eter !. "#ssell$ iGenetics% &opyright ' earson Ed#cation$ (nc.$p#)lishing as *en+amin mmings.

    24

  • 7/25/2019 Investigatory Project Biology

    25/45

    #loning a -ene Using 4acteria

    5tep ;7 'nsert the BGH-ene into the 4acterial"lasmid

    The acterial plasmid is also cut with the restrictionen*yme, leaving stic!y ends

    A plasmid is a small circular DNA that is separate fromthe acterial genome

    5tic!y ends of the cut 4-8 DNA attach ycomplementary ase pairing to the stic!y ends of

    the cut plasmid DNA

    This is now recominant DNA

    25

  • 7/25/2019 Investigatory Project Biology

    26/45

    Fig. 16.,&leavage of DNA )y the restriction enzyme Eco"(

    26

  • 7/25/2019 Investigatory Project Biology

    27/45

    eter !. "#ssell$ iGenetics% &opyright ' earson Ed#cation$ (nc.$ p#)lishing as *en+amin mmings.

    27

  • 7/25/2019 Investigatory Project Biology

    28/45

    #loning a -ene Using 4acteria

    5tep C7 'nsert the $ecominant "lasmid into a 4acterial #ell The recominant plasmid containing the r4-8 is then placed into

    acterial cells

    5tep

  • 7/25/2019 Investigatory Project Biology

    29/45

    TaDa $ecominant proteins

    C+#"#$ / G%#% U)"#$ B/*(%&"/

    8ow can acteria produce the cow 4-8 protein:

    This wor!s ecause acteria use the same geneticcode as cows %and all living things&

    2ther proteins are made in this way6

    'nsulin for diaetics

    #lotting factors for hemophiliacs

    #ancer treatment drugs

    #loning a -ene Using 4acteria

    29

  • 7/25/2019 Investigatory Project Biology

    30/45

    Using the lacEgene as areporter of

    geneepression

    $eporter gene proteinencoding genewhose

    epression in

    30

  • 7/25/2019 Investigatory Project Biology

    31/45

    the cell is quantifiale y techniques of proteindetection7

    Fusion of reporter gene to cis acting %DNA&regulatory regions %li!e promoters& allowsassessment gene activity y monitoring amount of

    reporter gene product

    $%&@

    Fusion used to perform genetic studies of the regulatory regionof gene G

    31

  • 7/25/2019 Investigatory Project Biology

    32/45

    $%&@H

    'ig( )*()+ a

    #loning is -enetic (ngineering

    32

  • 7/25/2019 Investigatory Project Biology

    33/45

    C+#"#$is the ma!ing of entire organisms usinggenetic engineering

    8as een done in cattle, goats, mice, cats, pigs,raits, and sheep

    8as never een done in humans

    33

  • 7/25/2019 Investigatory Project Biology

    34/45

    #loning is -enetic (ngineering

    Dolly the sheep was the first animal to e cloned

    The DNA %all CI chromosomes& from an adult sheep mammary glandwere fused with an unfertili*ed egg cell without any DNA inside

    The treated egg was placed in the uterus of an adult sheep that had received hormone treatments to support pregnancy

    There were @JJ failures efore this #5*%/& (&/#)!%&techniquesucceeded

    Dolly was successfully created in ?HHJ

    #loning is-enetic

    34

  • 7/25/2019 Investigatory Project Biology

    35/45

    35

  • 7/25/2019 Investigatory Project Biology

    36/45

    (ngineering

    36

  • 7/25/2019 Investigatory Project Biology

    37/45

    #loning is -enetic (ngineering

    Dolly was put to sleep at the age of I in @KK; 4ecause of health prolems

    5he was suffering from arthritis and a progressive lung disease

    These are usually only seen in old sheep

    #loned animals seem to age prematurely and show signs ofother health prolems that are normally associated withaging

    8ypothesis6

    (gg and 5perm DNA is reprogrammedB and does not reflect the age of theparents

    Adult donor DNA is not reprogrammedB in the egg and reflects the age of thedonor

    37

  • 7/25/2019 Investigatory Project Biology

    38/45

    Using adult DNA to create new organisms results in organisms that remain atthe age of their donor DNA at irth

    Therapeutic #loning Not cloning of entire organisms, ut cloning of specific

    tissues or cells

    "ancreatic cells to produce insulin in diaetics

    5pinal cord cells in paralli*ed patients

    S(% *%)are induced in the laoratory to turn into specifictissue cells

    +hat are 5tem #ells:

    #ells that can e induced to turn into every type of cell in the humanody

    +here are 5tem #ells found: 'n emryos

    38

  • 7/25/2019 Investigatory Project Biology

    39/45

    The original fertili*ed egg grows into an entire human eing7 Thesecells can ma!e every cell type

    (mryonic 5tem #ells

    39

  • 7/25/2019 Investigatory Project Biology

    40/45

    http699sps7!?@7ar7us9massengale9images9ivf7>pg

    5tem #ells40

  • 7/25/2019 Investigatory Project Biology

    41/45

    8ow do scientists acquire 5tem #ells:

    8uman eggs are fertili*ed y human sperm in vitro

    %in a test tue&

    Fertili*ed egg grows and divides y mitosis to anemryo which is >ust a all of cells at this point

    These cells can now e fro*en as stem cells for research

    This is the same process that is used y couplesthat cannot conceive a ay

    #alled in vitro fertili*ation %'/F&

    '/F

    41

  • 7/25/2019 Investigatory Project Biology

    42/45

    in vitro fertili*ation %'/F&

    The all of cells is implanted into the female)s uterus

    5he has een treated with hormones to simulate pregnancy to accept

    the emryohttp699pregnancyanday7com9pregnancy9ay9Facing1infertility1

  • 7/25/2019 Investigatory Project Biology

    43/45

    #ouples using '/F generally generate ?ust grow as individual stem cells

    $esearchers can grow millions and millions of these in the la to perform studies thatmay someday save lives and cure diseases

    43

  • 7/25/2019 Investigatory Project Biology

    44/45

    #ell 5tage(mryonic

    5tem #ells

    44

  • 7/25/2019 Investigatory Project Biology

    45/45

    5tem #ells

    The use of emryonic stem cells in research fuels aheated national deate

    3ostly ecause of scientific ignorance

    (mryonic stem cells are valued y researchers

    ecause they are (+("'+(%#(, or ale to ecome anyother cell

    +ith increased study, these could potentially treat or cureany type of disease and cancer

    'n @KK?, "resident 4ush anned federal funding forreaching using emryonic stem cells

    4ecause he never too! 4io ;IK

    %or any iology for that matter&45