‹#›
=
Ústav biologie a
lékařské genetiky 1.LF UK a VFN, Praha=
Molekulární genetika II=
=
![](3D"slide0022_image002.gif")
![](3D"slide0022_image004.gif")
![](3D"slide0022_image005.gif")
zimní
semestr
4.
výukový týden
(27.10. – 31.10.2008)&#=
13;
![](3D"slide0043_image006.gif")
DNA diagnostika
prenatáln&ia=
cute;
presymptomatick&aac=
ute;
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
![](3D"slide0043_image007.gif")
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
Potvrzení
diagnózy
Diagnostika
heterozygotů (přenašečů)
![](3D"slide0043_image008.gif")
![](3D"slide0043_image009.gif")
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
dělení podle cíle v genomu :
![](3D"slide0043_image010.gif")
P"=
5;ímá
diagnostika
Stanovení alely (mutace)
způsobující onemocnění
<=
/span>
=
Nep&#=
345;ímá
diagnostika
Stanovení
alely (markeru) v těsné vazbě s ge=
nem
nesoucím neznámou mutaci
způsobující onemocněn&=
iacute;
<=
/span>
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
![](3D"slide0044_image011.gif")
Přímá DNA diagnostika<=
/b>
=
hledáme
alelu (variantu), která zodpovídá za daný =
fenotyp
(onemocnění) v rodině
=
![](3D"slide0044_image012.gif")
u rodiče zjišt&#=
283;na
mutace v genu
![](3D"slide0044_image013.gif")
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
=
![](3D"slide0044_image014.gif")
u plodu
hledána tato mutace
Napříkl=
ad
– AD onemocnění, prenatální dg.:
![](3D"slide0044_image015.gif")
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
![](3D"slide0045_image016.gif")
=
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
Nepřímá DNA diagnostik=
a
![](3D"slide0045_image017.gif")
=
Nemusíme
vědět jaká mutace genu je zodpovědná za
onemocnění v rodině, postač&ia=
cute;
víme-li o který gen se jedná
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
=
při
diagnostice je využita genetická vazba mezi známý=
;m markerem,
který leží blízko genu a neznámo=
u mutac&iac=
ute; uvnitř genu.
Která alela markeru kosegreguje s
onemocněním? Má tuto alelu vyšetřovaná=
osoba?
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
=
=
mutace
marker
gen
chromozom
![](3D"slide0045_image019.gif")
u rodičů
zjištěny alely markeru
obvykle se sleduje
více markerů
![](3D"slide0045_image020.gif")
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
=
![](3D"slide0045_image021.gif")
u potom=
ků
zjištěny alely markeru
![](3D"slide0041_image022.gif")
Nepřím&aacut=
e;
diagnostika
=
neznáme
jaká mutace genu je zodpovědná za onemocnění=
; v rodině, pouze
víme o který gen se jedná
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
=
při
diagnostice je využita genetická vazba mezi známý=
;m markerem,
který leží blízko genu a neznámo=
u mutac&iac=
ute; uvnitř genu.
Která alela markeru kosegreguje s
onemocněním? Má jí vyšetřovaná
osoba?
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
HLAVN&I=
acute;
ÚSKALÍ nepřímé diagnostiky: U
různých jedinců jsou ve vazbě s mutovanou ale=
lou
(která způsobuje onemocnění) různé alely markeru.
Naopak
stejná alela markeru může jednou být ve vazbě=
s mutovanou alelou genu,
podruhé se standardní alelou genu.
Tedy
vyšetření může být neinformativní
nebo jen částečně =
informativní.
=
Vedlejší
potíž: možnost rekombinace mezi markerem a genem. Jej&iac=
ute;
pravděpodobnost je malá a celkem přesně
vyčíslitelná
![](3D"slide0042_image023.gif")
Variabilita na úr=
ovni
DNA
v
populaci se určité místo sekvence DNA vyskytuje v
různých formách -
alely
=
variabilní
místo se může nalézat jak uvnitř genů (v
exonech - tzv. kódující sekvence, v intron=
ech
- tzv. nekódující sekvence), tak mimo geny (v=
82;dy
tzv. nekódující sekvence)
dělení dle frekvence alel:
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
![](3D"slide0042_image024.gif")
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
![](3D"slide0042_image025.gif")
polym=
orfizmus
frekvence
„vzácnější“ alely je v populaci =
;
> 1%
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
̶=
2;mutace“,
„vzácná alela“
![](3D"slide0042_image026.gif")
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
frekv=
ence v
populaci < <=
/span>1%
dělení dle funkčního vlivu:
funk&=
#269;ní
varianta
alela=
je
spojena se změnou fenotypu
=
tich&=
aacute;
varianta
alela
nemá funkční dopad a ve fenotypu se
neprojeví
=
![](3D"slide0046_image027.gif")
Variabilita na úrovni DNA
dě=
lení
podle typu:
SNP
Single
nucleotide polymorphism =3D polymorfizmus jednoho neukleotidu
=
tande=
mové
repetice
mikrosatelity, také zvané např. STR =3D=
short
tandem repeats, minisatelity
<=
span
lang=3DCS style=3D'color:green;mso-special-format:lastCR;display:none'>=
3;
Str=
ukturní
polymorfizmy
Inzerce,
delece, inverze (1 bp – chromozom)
=
Dup=
likace, „low copy repeats“
Vlastně druh tandemových repetic, ale značně
odlišný co do techniky detekce a funkčního
významu Např. FCGR3B a riziko glomerulonefritidy, <=
span
lang=3DCS style=3D'font-size:250%;color:#404040'>RhD<=
span
lang=3DCS style=3D'font-size:250%;color:#404040'> a RhCE
=
SNP – single nucleotide
polymorphisms
Jednonukleotidové polymorfizmy=
TAG=
CCATCGGTA N GTACTCAATGATCA
A
![](3D"slide0036_image028.gif")
=
T
G
![](3D"slide0036_image029.gif")
C
![](3D"slide0036_image033.gif")
SNP
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
C
/ A
![](3D"slide0036_image037.gif")
1
V 99.9% sekvence DNA se lidé od sebe vzájemně
neliší. Ze zbývajícího 0.1% rozdílu
tvoří SNP přes 80%. Možnost typizace až miliónu SNP v jednom vzorku
pomocí DNA čipů by měla usnadnit identifikaci alel,
zodpovědných za řadu prevalentních
onemocnění.
![](3D"slide0036_image038.gif")
=
=
=
=
Možnosti stanovení SNP:
SNP ja=
ko
Polymorfismus délky restrikčních fragmentů
RFLP=
230;restriction
fragment length polymorphism
Restrikční
endonukleázy – bakteriální enzymy
štěpící DNA v místě specifické =
=
sekvence, chrání bakte=
rie
před infekcí bakteriofágem (a nebo taky ne)<=
/span>
5´=
; TAGCCATCGGTACGTACTCAATGATCA 3´
5´=
; TAGCCATCGGTAAGTACTCAATGATCA 3´
3&acut=
e;
ATCGGTAGCCATGCATGAGTTACTAGT 5´
3&acut=
e;
ATCGGTAGCCATTCATGAGTTACTAGT 3´
![](3D"slide0039_image039.gif")
RsaI
![](3D"slide0024_image040.gif")
=
Eco RI
Názvosloví
restrikčních endonukleáz (restriktáz)=
![](3D"slide0024_image041.gif")
Třípísmenná zkr=
atka
organizmu
„Escherichia coli“ <= /b>
„Escherichia coli“ <= /b>
Bližší
rozlišovací označení původu
RI…enzym pocházejíc&iacut= e; z kmene RY13, determinovaný R plazmidem <= /b>
RI…enzym pocházejíc&iacut= e; z kmene RY13, determinovaný R plazmidem <= /b>
![](3D"slide0024_image043.gif")
http://rebase.n=
eb.com
5
![](3D"slide0002_image044.gif")
=
![](3D"slide0002_image045.gif")
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
![](3D"slide0002_image046.gif")
=
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
Taq I 5´T C G A3&acu=
te; …=
5´A C G G T C G
A A T T3´
Taq I
5´T C G A<=
span
lang=3DCS style=3D'font-size:233%;text-shadow:auto;position:relative;top:-=
.45em;
mso-text-raise:30%'>3´ … 5´A C G G T C G A A T T3´
3´A G C T5´ …=
3´T G C C A G C T T A=
A5´
Hae III
5´G G C C 3´
C
C G G
=
Bam HI &n=
bsp;
5´G <=
/span>| G A T C C3´
![](3D"slide0002_image047.gif")
![](3D"slide0002_image048.gif")
![](3D"slide0002_image049.gif")
![](3D"slide0002_image051.gif")
Každá restriktáza
rozeznává specifickou sekvenci =
6
Štěpení dsDNA
restrikční endonukleázou EcoRI
![3Decori](3D"slide0023_image052.gif")
7
![](3D"slide0003_image053.gif")
restrikční
místo=
Alu I<=
b>
AG / CT
Sau 3AI <=
/b>
/ GTAC
Msp I<=
b>
C / CGG
5´G.G.G.C.G.T.A.C.A.T.A.G.C.T.A.A.T.G.G.C.A.A.G.C.=
T.A.T.G.G.T.3=
´
Najděte
restrikční místa pro nabídnuté enzymy na
uvedeném vlákně:
![](3D"slide0003_image054.gif")
=
![](3D"slide0003_image055.gif")
![](3D"slide0003_image056.gif")
![](3D"slide0003_image057.gif")
![](3D"slide0003_image058.gif")
![](3D"slide0003_image059.gif")
![](3D"slide0003_image060.gif")
![](3D"slide0003_image061.gif")
![](3D"slide0003_image062.gif")
![](3D"slide0003_image063.gif")
=
Identifikace restrikčních
míst
![](3D"slide0025_image064.gif")
Najděte
restrikční místa pro nabídnuté enzymy na
uvedeném vlákně:
Identifikace restrikčních
míst
C / CGG
Msp I<=
b>
/ GTAC
Sau 3AI <=
/b>
AG / CT
Alu I<=
b>
restrikční
místo=
5´G.G.G.C.G.T.A.C.A.T.A.G.C.T.A.A.T.G.G.C.A.A.G.C.=
T.A.T.G.G.T.3=
´
![](3D"slide0025_image068.gif")
2x
1x=
3;
0
9
![](3D"slide0005_image069.gif")
Kterou z
nabízených restriktáz můžete použ&iacu=
te;t
:
a) pro linearizaci
dané cirkulární struktury
b) pro
vyštěpení takového úseku, aby obsahoval pol=
y-A
sekvenci.
G / GATCC
Bam HI=
CGAT / CG
Pvu I<=
b>
T / CGA
Taq I<=
b>
restrikční
místo=
![](3D"slide0005_image070.gif")
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
![](3D"slide0005_image071.gif")
![](3D"slide0005_image072.gif")
![](3D"slide0005_image073.gif")
![](3D"slide0005_image074.gif")
=
![](3D"slide0005_image075.gif")
![](3D"slide0005_image076.gif")
![](3D"slide0005_image077.gif")
![](3D"slide0005_image078.gif")
![](3D"slide0005_image079.gif")
Identifikace restrikčních
míst
![](3D"slide0005_image080.gif")
![](3D"slide0005_image081.gif")
10
![](3D"slide0010_image083.gif")
![](3D"slide0010_image084.gif")
Analýza Southernova blotu. &#=
13;
Popis metody<=
span
style=3D'font-size:300%;color:blue;mso-color-index:0;mso-special-format:la=
stCR;
display:none'>
![](3D"slide0010_image085.gif")
=
http://www.ac-creteil.fr/biotechnologies/doc_southernblotting.htm
11
Southern blot – animace
externí soubor
externí soubor
![](3D"slide0009_image086.gif")
![](3D"slide0009_image087.gif")
![](3D"slide0009_image088.gif")
![](3D"slide0009_image089.gif")
![](3D"slide0009_image090.gif")
DNA
Sonda
Zahřát&=
iacute;
(denaturace)
Ochlazení=
3;
(renaturace)
![](3D"slide0009_image091.gif")
![](3D"slide0009_image092.gif")
![](3D"slide0009_image093.gif")
![](3D"slide0009_image094.gif")
Southern blot – hybridizace sondy
12
![](3D"slide0011_image095.gif")
Úkol č. 4, str. 74 Gen G
![](3D"slide0011_image096.gif")
![](3D"slide0011_image097.gif")
![](3D"slide0011_image098.gif")
![](3D"slide0011_image099.gif")
![](3D"slide0011_image100.gif")
![](3D"slide0011_image101.gif")
![](3D"slide0011_image102.gif")
![](3D"slide0011_image103.gif")
![](3D"slide0011_image104.gif")
![](3D"slide0011_image105.gif")
![](3D"slide0011_image106.gif")
![](3D"slide0011_image107.gif")
![](3D"slide0011_image108.gif")
![](3D"slide0011_image109.gif")
![](3D"slide0011_image110.gif")
![](3D"slide0011_image111.gif")
![](3D"slide0011_image112.gif")
![](3D"slide0011_image113.gif")
![](3D"slide0011_image114.gif")
![](3D"slide0011_image115.gif")
![](3D"slide0011_image116.gif")
![](3D"slide0011_image117.gif")
![](3D"slide0011_image118.gif")
![](3D"slide0011_image119.gif")
![](3D"slide0011_image120.gif")
![](3D"slide0011_image121.gif")
![](3D"slide0011_image122.gif")
1 kb
5´
3´
![](3D"slide0011_image123.gif")
=
![](3D"slide0011_image124.gif")
![](3D"slide0011_image125.gif")
![](3D"slide0011_image126.gif")
![](3D"slide0011_image127.gif")
![](3D"slide0011_image128.gif")
Taq I
EcoR I=
*
![](3D"slide0011_image129.gif")
=
![](3D"slide0011_image130.gif")
sonda:
![](3D"slide0011_image131.gif")
=
cDNA
genomová
=
13
Úkol č. 4a, str. 74 Gen G – prosté
štěpení=
![](3D"slide0032_image158.gif")
1 kb
5´
3´
![](3D"slide0032_image159.gif")
=
![](3D"slide0032_image160.gif")
![](3D"slide0032_image161.gif")
Taq I
*
=
Výsledky:
a) na 4 nebo 5 fragmentů:
3kb, 8kb, 6kb/2kb+4kb, 7kb
![](3D"slide0032_image162.gif")
5´
3´
![](3D"slide0032_image163.gif")
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
![](3D"slide0032_image164.gif")
![](3D"slide0032_image165.gif")
![](3D"slide0032_image166.gif")
![](3D"slide0032_image167.gif")
![](3D"slide0032_image168.gif")
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
![](3D"slide0032_image169.gif")
=
![](3D"slide0032_image178.gif")
14
![](3D"slide0034_image179.gif")
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
=
=
=
Úkol č. 4b, c, str. 74 Gen G – Southern blot s cDNA sondou=
![](3D"slide0034_image206.gif")
1 kb
5´
3´
![](3D"slide0034_image207.gif")
=
![](3D"slide0034_image208.gif")
![](3D"slide0034_image209.gif")
Taq I
*
=
Výsledky:
![](3D"slide0034_image210.gif")
5´
3´
![](3D"slide0034_image211.gif")
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
![](3D"slide0034_image212.gif")
![](3D"slide0034_image213.gif")
![](3D"slide0034_image214.gif")
![](3D"slide0034_image215.gif")
![](3D"slide0034_image216.gif")
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
![](3D"slide0034_image217.gif")
=
b)&nb=
sp;
3
fragmenty: 3kb, 8kb, 7kb
c) ne
15
![](3D"slide0035_image227.gif")
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
Úkol č. 4d, str. 74 Gen G – Southern blot s genomickou =
=
; sondou&#=
13;
![](3D"slide0035_image254.gif")
1 kb
5´
3´
![](3D"slide0035_image255.gif")
=
![](3D"slide0035_image256.gif")
![](3D"slide0035_image257.gif")
Taq I
*
=
Výsledky:
![](3D"slide0035_image258.gif")
5´
3´
![](3D"slide0035_image259.gif")
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
![](3D"slide0035_image260.gif")
![](3D"slide0035_image261.gif")
![](3D"slide0035_image262.gif")
![](3D"slide0035_image263.gif")
![](3D"slide0035_image264.gif")
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
![](3D"slide0035_image265.gif")
=
d) Ano 4kb (alela
+) 6kb (alela -)
16
![](3D"slide0026_image275.gif")
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
=
![](3D"slide0026_image276.gif")
![](3D"slide0026_image277.gif")
![](3D"slide0026_image278.gif")
![](3D"slide0026_image279.gif")
![](3D"slide0026_image280.gif")
![](3D"slide0026_image281.gif")
![](3D"slide0026_image282.gif")
![](3D"slide0026_image283.gif")
![](3D"slide0026_image284.gif")
![](3D"slide0026_image285.gif")
![](3D"slide0026_image286.gif")
![](3D"slide0026_image287.gif")
![](3D"slide0026_image288.gif")
![](3D"slide0026_image289.gif")
![](3D"slide0026_image290.gif")
![](3D"slide0026_image291.gif")
=
M
![](3D"slide0026_image292.gif")
![](3D"slide0026_image293.gif")
![](3D"slide0026_image294.gif")
=
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
![](3D"slide0026_image295.gif")
![](3D"slide0026_image296.gif")
![](3D"slide0026_image297.gif")
![](3D"slide0026_image298.gif")
![](3D"slide0026_image299.gif")
![](3D"slide0026_image300.gif")
![](3D"slide0026_image301.gif")
![](3D"slide0026_image302.gif")
![](3D"slide0026_image303.gif")
![](3D"slide0026_image304.gif")
![](3D"slide0026_image305.gif")
![](3D"slide0026_image306.gif")
![](3D"slide0026_image307.gif")
![](3D"slide0026_image308.gif")
![](3D"slide0026_image309.gif")
=
P
![](3D"slide0026_image310.gif")
=
E1 – restrikční endonukleáza 1&=
#13;
=
E2 –
restrikční endonukleáza 2
M
– fragment maternálního chromosomu
![](3D"slide0026_image311.gif")
![](3D"slide0026_image312.gif")
------=_NextPart_01C945D6.54226D30
Content-Location: file:///C:/0579C638/MolGenII_v2008_soubory/slide0026.xml
Content-Transfer-Encoding: quoted-printable
Content-Type: text/xml; charset="utf-8"
P
– fragment paternálního chromosomu
¯ - místa
štěpení restriktázami (cílové
sekvence)
=
2cM
– vzdálenost sledovaným genem a místem, kde
hybridizuje sonda, resp. místem restrikce=
=
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
Mimogenová sonda
17
![](3D"slide0030_image313.gif")
=
M
=
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
![](3D"slide0030_image314.gif")
E1
![](3D"slide0030_image315.gif")
E1
![](3D"slide0030_image316.gif")
E1
![](3D"slide0030_image317.gif")
E1
![](3D"slide0030_image318.gif")
E1
![](3D"slide0030_image319.gif")
E1
![](3D"slide0030_image320.gif")
E1
![](3D"slide0030_image321.gif")
![](3D"slide0030_image322.gif")
![](3D"slide0030_image323.gif")
=
P
![](3D"slide0030_image324.gif")
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
Mimogenová sonda –
štěpení restriktázou E1
Ačkoli
restriktáza E1 bude štěpit DNA ve všech
restrikčních místech, na autoradiogramu detekujeme jenom fragmen=
ty, na
které hybridizovala sonda, a ty jsou u obou chromozómů
stejně dlouhé –
jedinec je homozyg=
ot v
délce restrikčních fragmentů.
![](3D"slide0030_image325.gif")
=
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
![](3D"slide0030_image326.gif")
E1
![](3D"slide0030_image327.gif")
E1
![](3D"slide0030_image328.gif")
E1
![](3D"slide0030_image329.gif")
E1
![](3D"slide0030_image330.gif")
E1
![](3D"slide0030_image331.gif")
E1
![](3D"slide0030_image332.gif")
E1
![](3D"slide0030_image333.gif")
![](3D"slide0030_image334.gif")
![](3D"slide0030_image335.gif")
![](3D"slide0030_image336.gif")
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
![](3D"slide0030_image337.gif")
![](3D"slide0030_image338.gif")
E1
=
autoradiogram=
18
![](3D"slide0031_image339.gif")
=
M
=
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
![](3D"slide0031_image340.gif")
E2
![](3D"slide0031_image341.gif")
E2
![](3D"slide0031_image342.gif")
E2
![](3D"slide0031_image343.gif")
E2
![](3D"slide0031_image344.gif")
E2
![](3D"slide0031_image345.gif")
![](3D"slide0031_image346.gif")
![](3D"slide0031_image347.gif")
=
P
![](3D"slide0031_image348.gif")
E2
![](3D"slide0031_image349.gif")
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
Mimogenová sonda –
štěpení restriktázou E2
Ačkoli
restriktáza E2 bude štěpit DNA ve všech
restrikčních místech, na autoradiogramu detekujeme jenom fragmen=
ty, na
které hybridizovala sonda, a fragment vyštěpený z
maternálního chromozómu je kratší –
jedinec je heteroz=
ygot v
délce restrikčních fragmentů.
![](3D"slide0031_image350.gif")
=
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
![](3D"slide0031_image351.gif")
E2
![](3D"slide0031_image352.gif")
E2
![](3D"slide0031_image353.gif")
E2
![](3D"slide0031_image354.gif")
E2
![](3D"slide0031_image355.gif")
E2
![](3D"slide0031_image356.gif")
![](3D"slide0031_image357.gif")
![](3D"slide0031_image358.gif")
![](3D"slide0031_image359.gif")
E2
![](3D"slide0031_image360.gif")
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
![](3D"slide0031_image361.gif")
![](3D"slide0031_image362.gif")
![](3D"slide0031_image363.gif")
E1
E2
=
autoradiogram=
E2
=
19
![](3D"slide0014_image364.gif")
![](3D"slide0014_image365.gif")
![](3D"slide0014_image366.gif")
=
![](3D"slide0014_image367.gif")
=
![](3D"slide0014_image368.gif")
=
![](3D"slide0014_image369.gif")
![](3D"slide0014_image370.gif")
![](3D"slide0014_image371.gif")
![](3D"slide0014_image372.gif")
![](3D"slide0014_image373.gif")
![](3D"slide0014_image374.gif")
![](3D"slide0014_image375.gif")
![](3D"slide0014_image376.gif")
![](3D"slide0014_image377.gif")
![](3D"slide0014_image378.gif")
=
1
Rodina
je tzv. neinformativní. Všichni jsou homozygoti v
délce restrikčních fragmentů . (Rodiče heterozygoti ve sledovaném genu). Nevíme však, který
fragment 1 p ředali, zda s mutovan=
ou
nebo zdravou alelou.
![](3D"slide0014_image379.gif")
=
![](3D"slide0014_image380.gif")
![](3D"slide0014_image381.gif")
=
![](3D"slide0014_image382.gif")
![](3D"slide0014_image383.gif")
=
![](3D"slide0014_image384.gif")
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
![](3D"slide0014_image385.gif")
![](3D"slide0014_image386.gif")
![](3D"slide0014_image387.gif")
![](3D"slide0014_image388.gif")
![](3D"slide0014_image389.gif")
=
1
![](3D"slide0014_image390.gif")
=
2
Rodina
je jen částeč ně informativní,
proto=
že nelze rozhodnout, =
se
kterým fragmentem je ve vazbě mutovan alela u otce a matk=
y.
Nicmé<=
span
lang=3DCS style=3D'font-family:Arial;mso-ascii-font-family:Arial;mso-bidi-=
font-family:
"Times New Roman";font-size:250%;text-shadow:auto'>ně , druhý potom=
ek
je homozygot AA nebo aa.
20
![](3D"slide0015_image391.gif")
![](3D"slide0015_image392.gif")
![](3D"slide0015_image393.gif")
=
![](3D"slide0015_image394.gif")
![](3D"slide0015_image395.gif")
=
![](3D"slide0015_image396.gif")
![](3D"slide0015_image397.gif")
![](3D"slide0015_image398.gif")
![](3D"slide0015_image399.gif")
![](3D"slide0015_image400.gif")
![](3D"slide0015_image401.gif")
=
1
![](3D"slide0015_image402.gif")
=
2
=
Z otcovy
strany fragment 2 ve vazbě s mutovanou alelou, protože postižený syn m&aac=
ute;
fragment 2 od otce. Matka je totiž homozygot (1,1) a tudíž p ředává
potomkům pouze fragmenty 1.
Nelze ale rozhodnout, zda druhému dítěti předala fragment se
zdravou nebo mutovanou alelou. Druhé dítě bude bu<=
span
lang=3DCS style=3D'font-size:250%;text-shadow:auto'>ď v 50% postiženo nebo v 50% p=
řenašeč.
![](3D"slide0015_image403.gif")
=
=
Zde
nebude analýza úspěšná,
chybí-li DNA zemřelého syna. Riziko 25%. Rodiče jsou heterozygoti
– měli posti<=
span
lang=3DCS style=3D'font-size:250%;text-shadow:auto'>žené dí=
tě AR chorobou
21
![](3D"slide0016_image404.gif")
![](3D"slide0016_image405.gif")
![](3D"slide0016_image406.gif")
![](3D"slide0016_image407.gif")
=
![](3D"slide0016_image408.gif")
![](3D"slide0016_image409.gif")
![](3D"slide0016_image410.gif")
![](3D"slide0016_image411.gif")
![](3D"slide0016_image412.gif")
![](3D"slide0016_image413.gif")
![](3D"slide0016_image414.gif")
=
1
![](3D"slide0016_image415.gif")
![](3D"slide0016_image416.gif")
=
2
Postižený syn je
homozygot v délce restrikč ních fragmentů <=
span
style=3D'position:absolute;top:61.5%;left:6.55%;width:40.26%;height:4.5%'>=
(1,1). Z toho plyne,=
že =
=
fragmenty 1 jsou u o=
bou =
rodičů s mutovanou =
alelou. Jestliže druhé
dít ě je =
homozygot (2,2),
znamená to, že bude zdravý
homozygot ve sledovaném genu.=
![](3D"slide0016_image417.gif")
=
![](3D"slide0016_image418.gif")
![](3D"slide0016_image419.gif")
![](3D"slide0016_image420.gif")
![](3D"slide0016_image421.gif")
![](3D"slide0016_image422.gif")
![](3D"slide0016_image423.gif")
![](3D"slide0016_image424.gif")
=
![](3D"slide0016_image425.gif")
=
![](3D"slide0016_image426.gif")
![](3D"slide0016_image427.gif")
![](3D"slide0016_image428.gif")
![](3D"slide0016_image429.gif")
![](3D"slide0016_image430.gif")
![](3D"slide0016_image431.gif")
=
1
=
2
Otec
(1,1), matka (1,2), postižený syn (1,2) musí
mít fragment 1 od otce a od matky fragment 2,
který je ve vazbě s mutovanou
alelou. Druhé dít ě (1,1) má od m=
atky
fragment 1, který nese zdravou alelu a=
od
otce také fragment 1, ale nevíme, zda s =
mutovanou nebo zdrav=
ou
alelou. Prognóza: 50% heterozygot, 50% zdravý homozy=
got
ve sledovaném genu - fenotypov=
ě zdravé.<=
/span>
![](3D"slide0016_image432.gif")
=
![](3D"slide0016_image433.gif")
![](3D"slide0016_image434.gif")
![](3D"slide0016_image435.gif")
22
![](3D"slide0017_image436.gif")
Jsou jedinci II/2 a=
II/3
heterozygoti pro mutovanou alelu sledovaného AR
onemocnění? (vazba úplná)
![](3D"slide0017_image437.gif")
![](3D"slide0017_image438.gif")
![](3D"slide0017_image439.gif")
![](3D"slide0017_image440.gif")
![](3D"slide0017_image441.gif")
![](3D"slide0017_image442.gif")
![](3D"slide0017_image443.gif")
1
![](3D"slide0017_image444.gif")
![](3D"slide0017_image445.gif")
![](3D"slide0017_image446.gif")
2
![](3D"slide0017_image447.gif")
![](3D"slide0017_image448.gif")
![](3D"slide0017_image449.gif")
I
II
![](3D"slide0017_image450.gif")
=
1
![](3D"slide0017_image451.gif")
=
1
![](3D"slide0017_image452.gif")
=
2
![](3D"slide0017_image453.gif")
=
2
3
aa
AA
![](3D"slide0017_image454.gif")
a
![](3D"slide0017_image455.gif")
a
![](3D"slide0017_image456.gif")
A
![](3D"slide0017_image457.gif")
A
Úkol č. 1a, str. 80
23
![](3D"slide0018_image458.gif")
Jsou jedinci II/2 a=
II/3
heterozygoti pro mutovanou alelu sledovaného AR
onemocnění? (vazba úplná)
![](3D"slide0018_image459.gif")
![](3D"slide0018_image460.gif")
![](3D"slide0018_image461.gif")
![](3D"slide0018_image462.gif")
![](3D"slide0018_image463.gif")
![](3D"slide0018_image464.gif")
![](3D"slide0018_image465.gif")
1
![](3D"slide0018_image466.gif")
![](3D"slide0018_image467.gif")
![](3D"slide0018_image468.gif")
2
![](3D"slide0018_image469.gif")
![](3D"slide0018_image470.gif")
![](3D"slide0018_image471.gif")
I
II
![](3D"slide0018_image472.gif")
=
1
![](3D"slide0018_image473.gif")
=
1
![](3D"slide0018_image474.gif")
=
2
![](3D"slide0018_image475.gif")
=
2
3
aa
AA
![](3D"slide0018_image476.gif")
a
=
![](3D"slide0018_image477.gif")
a
![](3D"slide0018_image478.gif")
A
=
![](3D"slide0018_image479.gif")
A
II/2 ne
II/3 ano<=
span
style=3D'font-size:300%;text-shadow:auto;mso-special-format:lastCR;display=
:none'>
Úkol č. 1a, str. 80
24
![](3D"slide0019_image480.gif")
Úkol č. 1b, str. 81
![](3D"slide0019_image481.gif")
![](3D"slide0019_image482.gif")
![](3D"slide0019_image483.gif")
![](3D"slide0019_image484.gif")
![](3D"slide0019_image485.gif")
=
1
![](3D"slide0019_image486.gif")
![](3D"slide0019_image487.gif")
=
2
![](3D"slide0019_image488.gif")
![](3D"slide0019_image489.gif")
![](3D"slide0019_image490.gif")
=
I
=
II
![](3D"slide0019_image491.gif")
=
![](3D"slide0019_image492.gif")
=
![](3D"slide0019_image493.gif")
=
![](3D"slide0019_image494.gif")
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
=
Jsou
jedinci II/2 a II/3 heterozygoti pro mutovanou alelu sledovan&eacu=
te;ho AR
onemocnění? (vazba úplná)
![](3D"slide0019_image495.gif")
II/2 AA nebo Aa
II/3 AA nebo Aa=
Vyšetření je
neúsp ěení
heterozygocie
jedinců II/2 a II/3).
![](3D"slide0019_image496.gif")
Aa
Aa
a
a
25
![](3D"slide0020_image497.gif")
Úkol č. 1c, str. 81
<=
/div>
![](3D"slide0020_image498.gif")
![](3D"slide0020_image499.gif")
![](3D"slide0020_image500.gif")
![](3D"slide0020_image501.gif")
![](3D"slide0020_image502.gif")
![](3D"slide0020_image503.gif")
![](3D"slide0020_image504.gif")
=
1
![](3D"slide0020_image505.gif")
=
2
![](3D"slide0020_image506.gif")
![](3D"slide0020_image507.gif")
![](3D"slide0020_image508.gif")
=
I
=
II
![](3D"slide0020_image509.gif")
=
![](3D"slide0020_image510.gif")
2
![](3D"slide0020_image511.gif")
![](3D"slide0020_image512.gif")
X+Y
X+Xh=
XhY
X+Y
![](3D"slide0020_image513.gif")
Xh
![](3D"slide0020_image514.gif")
X+
X+
![](3D"slide0020_image515.gif")
X+
II/3 ano<=
span
style=3D'font-size:350%;mso-special-format:lastCR;display:none'>
![](3D"slide0020_image516.gif")
Xh
Xh
26
![](3D"slide0021_image517.gif")
![](3D"slide0021_image518.gif")
![](3D"slide0021_image519.gif")
![](3D"slide0021_image520.gif")
![](3D"slide0021_image521.gif")
![](3D"slide0021_image522.gif")
=
1
![](3D"slide0021_image523.gif")
=
2
![](3D"slide0021_image524.gif")
![](3D"slide0021_image525.gif")
![](3D"slide0021_image526.gif")
=
I
=
II
![](3D"slide0021_image527.gif")
=
1
![](3D"slide0021_image528.gif")
=
2
![](3D"slide0021_image529.gif")
3
=
2
<=
span
lang=3DCS style=3D'mso-special-format:lastCR;display:none'>
1
Úkol č. 1d, str. 81
II/3 ne
![](3D"slide0021_image530.gif")
Xh
Xh
X+
X+
X+
X+
X+
27
Klepnutím lze upravit styly předlohy textu.
=
Druhá úroveň
Třetí úroveň
Čtvrtá úroveň
Pátá úroveň
‹#›
10<=
/div>
11<=
/div>
' + ENDSHOW_MESG + '
![](3D"notes_flag.gif")
Po=
známky![](3D"buttons.gif")
![](3D"slide0022_image001.png"){kind=link}
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