of λDNA Internship Epigenetics Dam Dam methylase CpG CpG methylase ClaI restriction enzyme MboI restriction enzyme SAM Co factor SAM Definitions ID: 930888
Download Presentation The PPT/PDF document "DNA- Methylation and Restriction" is the property of its rightful owner. Permission is granted to download and print the materials on this web site for personal, non-commercial use only, and to display it on your personal computer provided you do not modify the materials and that you retain all copyright notices contained in the materials. By downloading content from our website, you accept the terms of this agreement.
Slide1
DNA-
Methylation
and
Restriction
of
λ-DNA
Internship
Epigenetics
Slide2Dam
Dam
methylase
CpG
CpG methylaseClaI restriction enzyme
MboI
restriction enzyme
SAM
Co
factorSAMDefinitions
Slide35‘ … ACAGATCACAGATGACGATGACG
AT … 3‘
Sequence for Damto recognize
Enzyme:Dam methylasemethyl group
Sequence
for CpGto recognizeCo
factorSAM
SAM
Dam
Methylation
of λ-DNA with Dam
Slide45‘ … ACAGATCACAGATGACGATGACG
AT … 3‘
Sequence for Damto recognize
Sequence
for CpGto recognize
SAM
Dam
Methylation
of
λ-DNA with Dam
Slide55‘ … ACA
GATCACAGATGACGATGACG
AT … 3‘Methylation of λ-DNA with Dam
Sequence for Damto
recognize
Sequence
for CpGto recognize
Dam
Slide6Sequence
for Damto recognize
Enzyme:CpG methylasemethyl group
Sequence for CpGto recognizeCo
factor:SAM
SAM
CpG
5‘ … ACA
GATC
ACAGATGACGATGACGAT … 3‘Methylation of λ-DNA with CpG
Slide75‘ … ACA
GATC
ACAGATGACGATGACGAT … 3‘
CpG
Sequence for Damto recognize
Sequence for CpGto recognize
SAM
Methylation
of λ-DNA with CpG
Slide85‘ … ACA
GATC
ACAGATGACGATGACGAT … 3‘
CpG
Sequence for Damto recognize
Sequence for CpGto recognize
Methylation
of
λ-DNA with CpG
Slide9Restriction
of
Dam-methylated and non-methylated λ-DNA with
MboI Dam-methylated
Non-methylated5‘ … ACAGATCACAGATGACGATGA
CGAT … 3‘
Sequence
for MboIto
recognize and cutRestriction enzyme MboIMethyl group from DamRestriction enzyme MboI
GATCACAGATGACGATGACGAT … 3‘
5‘ … ACA
Slide10Restriction
of CpG-methylated and non-methylated
λ-DNA with ClaI
CpG-methylatedNicht methyliert5‘ … ACAACATCGAT
AATGAATCAT … 3‘
Sequence for ClaIto recognize and cut
Restriction
enzyme
ClaIMethyl group from CpG5‘ … ACAACATSequence for ClaIto recognize and cutCGATAATGAATCAT … 3‘Restriktionsenzym ClaI
Slide11Regular Case of a Restriction5‘ … ACA
CpG-methylated λ-DNA, cutted with MboI
Sequence
for CpGto recognize
Sequence for MboIto recognize and cut
GATC
ACAGATGA
CG
ATGA
CGAT … 3‘CpG methylated: Restriction enzyme MboI
Slide12CpG
methylated
5‘
… AA
GATCGCAGATGACGATGACGAT
… 3‘
Exception
for
a RestrictionCpG-methylated λ-DNA, cutted with MboI Sequence for CpGto recognizeSequence for MboIto recognize and cut
: Restriktionsenzym
MboI
Slide13Regular Case of a Restriction
5‘ … AC
GATCACAGATATDam-methylated λ-DNA, cutted
with ClaI CGATGAT … 3‘
Dam methylated
Sequence
for Damto recognize
Sequence
for
ClaIto recognize and cutClaI: Restriction enzyme ClaI
Slide14Exception
for a Restriction
5‘ … AA
GATCACAGATATCGATCAT … 3‘
Dam-methylated λ-DNA, cutted with ClaI
Dam methylated
Sequence
for
Damto recognize
Sequence for ClaIto recognize and cutClaI: Restriction enzyme ClaI
Slide15λ-DNA
= non-methylated Lambda-DNA λ-DNA MboI= non-methylated Lambda-DNA cutted with MboIλ-DNA
/Dam MboI= Dam-methylated Lambda-DNA cutted with MboI Proof of
λ-DNA fragments cutted with MboI
λ
-DNA
/Dam
MboI
λ
-DNA
MboI
λ
-
DNA
Slide16λ
-DNA= non-methylated Lambda-DNA λ-DNA ClaI= non-methylated Lambda-DNA cutted with ClaI
λ-DNA/CpG ClaI= CpG-methylated Lambda-DNA cutted with ClaI
Proof of λ-DNA fragments cutted with ClaI
λ
-DNA
/CpG
ClaI
λ
-DNA
ClaI
λ
-
DNA