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THS NGUYEN KHANH LINH

BAI GIANG
CHNG 1
A. Acid deoxyribonucleic
I. Cau truc DNA
1. Cau tao va cau truc DNA
a. Cau tao
- Moi nucleotide gom 1 base nit ( purin A,G hay pirimidin T,
C) + duong deoxyribose( khong co nhom OH o 2`-carbon
nh ng ribose) + goc phosphat
- 4 nucleotide: dATP (deoxyadenosine-triphosphat), dGTP
(deoxyguanosine-triphosphat), dCTP (deoxycytidine-
triphosphat), dTTP (deoxythymidine-triphosphat)
-Co huong 5` - 3`
- Phan biet nucleoside
- Cac base nit 2 hai mach lien ket hydro (A=T, G=C)
b. Cau truc xoan kep
- Mot vong xoan kep dai = 3.4nm, khoang 10.4 bp
thay oi
- ng knh vong xoan=2nm (2 pirimidin: ln, 2 purin:
mat on nh chuoi xoan kep)
- Khoang cach gia 2 cap base lien tiep= 0.34nm
1953, James Watson va
Francis Crick a a ra mo
hnh xoan kep cua DNA
V moi cap base c nh v
1 goc tren truc xoan helix
nen nhn tong the mo hnh
DNA nh mot cau thang
xoan oc
Doc theo phan t DNA, co 2
ranh. Ranh ln rong va sau,
ranh nho nong va can (la ni
protein se tng tac vi cac
base)
Cac loai lien ket khong hoa tr gop phan tao tnh on nh
cua chuoi xoan kep DNA:
Tng tac k nc
Lien ket hydro
Lien ket Van der waal
Tng tac tnh ien: be mat ben ngoai cua DNA ( do
khung ng - phosphat) co nhom phosphat tch ien
am. Luc y gia cac nhom phosphat ke can se c han
che nh pt hoa tr 2 nh Mg
2+
va histon, giup on nh
DNA
2.Cau truc sieu xoan
Xoan am: xoan phai, DNA xoan len chnh no
Xoan dng: xoan trai, thng gap khi DNA xoan lay
protein
Kiem soat sieu xoan: enzyme topoisomerase ( I, II)
oi vi mach DNA vong: khi sao chep, enzyme
topoisomerase
(Enzyme gyrase E.coli) se cat mach
II. Nhiem sac the va nhiem sac chat
1.Prokaryote
V du E.coli: DNA kep tao cau truc mach vong, 4.6Mb
(10
6
base), 4300 gen, 9 trieu nucleotide
NST mach vong vi khuan c nen chat tao cau truc co kch
thc 1mx2m
NST E.coli dang sieu xoan, gan ket vi protein loi, nam t
do trong te bao chat, goi chung la nucleoid
o nen: do HU (40 v tr gan), cac polyamine ( polycationic
pt nh spermidine, spermine)
2. Eukaryote
* Protein histon chia thanh 5 lp: H1,
H2A, H2B, H3, H4
* DNA n mach thang, lk protein
histon, protein phi histon, RNA va
polyamine
* Nucleosome: 2 ban sao cua H2A, H2B,
H3, H4, 140bp, DNA linker ( 60bp), 1 pt H1
Cac nucleosome quan thanh oan
ng knh 30nm( 30nm solenoid),
200nm
oan nay bao gom nhieu loop (oan
30nm) quan quanh 1 phc protein loi
nuclear scaffold
Moi NST eu co centromer (vung giau AT, lk protein
khong histon e tao thanh kinetochore noi vi thoi vo sac)
va telomere ( giau CCCA, can viec mat thong tin di truyen
trong qua trnh sao chep)
Khi khong phan chia te bao, NST dang ni long
chromatin. dang long, quan sat vi anh ien t thay co
nhng cham nucleosome
Phan biet:
* Heterochromatin: khong c phien ma.
* Euchromatin: se nen chat metaphase, cha phan
ln thong tin di truyen
Eukaryote, ti the va luc lap ?
II. Cau truc gen
1. Prokaryote
Bo gen c nen, lien tuc, t oan khong giai ma, mach
vong
Qua trnh ieu hoa cac gen c thc hien nh co cau truc
operon ( gom cac gen lien ket vi nhau, c ieu hoa
chung nh 1 n v)
Prokaryote thng co them oan DNA nho, plasmid, thng
dang vong, co vi khuan va mot so eukaryote
2. Eukaryote
Khac biet: ve kch thc gen, kha nang giai ma t (khong
qua 1.5% ngi giai ma protein; khong co trnh t khi
s phien ma)
Bo gen mach thang
Phc tap, gen gom intron ( tach ra khoi hnRNA e tao
thanh mRNA trng thanh), exon ( oan phien ma, ra
nhan, dch ma)
Y ngha: vai tro tien hoa, lien quan en cac domain
protein
ngi: 1.5% genome la exon, 23.5% genome la intron,
tRNA gen, rRNA gen. Moi bo 23 NST nhan t cha hay
me cha khoang 6.10
9
nucleotide hay 3.10
9
bp
Cac loai trnh t DNA:
Ho gen: cac gen trong cung 1 ho thng giai ma cho cac protein co
cung chc nang chuyen biet nhng vao cac thi iem khac nhau
trong qua trnh song, cac protein o gl isoform, c phan biet bang
cach them k hieu Latinh sau ten protein nh globin , globin
Satelitte DNA:Con khoang 45% genome Eukaryote cha c
hieu ro, co cac trnh t DNA nh nhau c lap i lap lai hn 1trieu
lan tren toan bo genome, thng vai tram cap base, tap trung
centromere
Minisatelitte ( 25 cap base) thng vung telomere; microsatellite (
<4bp): c dung lam dau hieu chan oan benh di truyen v chung
thay oi tuy loai sinh vat
Tandem DNA: la cac oan DNA (phien ma ra RNA) co trnh t nh
nhau, lap i lap lai tren cung mot NST, gom rRNA gen (250ban/tb),
tRNA(50 ban/tb), protein histon
Transposon: la nhng oan DNA co the tach ra va chen vao bat ky
v tr nao tren toan bo genome
Retrotransposon: qua ban sao phien ma trung gian
RNA
Alu sequence ngi: 280bp, 500.000 copies
B. Acid ribonucleic
I. Cau tao
Khac DNA: ng ribose, cac base : A, U, G, C
Mach n, polymere, co hng, t tao cau truc bac 2
2`-OH cua ribose co the tao lien ket hydro vi phan t ke
can
II.Phan loai
1. Messenger RNA
Mang thong tin t DNA e tong hp Protein
Chiem khoang 5% tong so RNA te bao, kch thc thay
oi
(E.coli: 500-6000 nucleotid)
Cistron: la trnh t DNA cha thong tin di truyen cho 1
chuoi polypeptid va vai tn hieu e ribosome thc hien
chc nang
Prokaryote: Polycistronic, mRNA c dch ma trong
khi hay ngay sau khi ket thuc phien ma
Eukaryote: Monocistronic, mRNA ch c dch ma
sau khi c chnh sa (cat xen, gan uoi poly-A, gan
cap metyl o dau 5`)
2. Transfer RNA
15% RNA te bao, trung bnh 75 nucleotide
Cau truc bac 3 nh cloverleaf ( co 3 la)
au 3` lien ket CHT voi aa, anticodon loop voi codon
D loop (dihydrouridin), TC loop (thymine, pseudouridin,
cytosine), variable loop : vtro?
Variable loop: e phan loai tRNA
( a so 80% cha 4-5 nucleotid, con lai c 20 nucleotide)
-Mang nhieu loai base bo sung nh pseudouridine,
dihydrouridine. . .
- Co nhieu loai, tng ng vi cac loai acid amin
3. Ribosome RNA
Chiem en 80% tong so RNA cua te bao
Ve trnh t nucleotid khac nhau tuy loai nhng cau truc 3
chieu hau nh bao ton
Thanh phan tao nen ribosome
4. Heterogeneous nuclear RNA (hnRNA)
La ban phien ma au tien, la tien than cua mRNA
5. Small nuclear RNA (snRNA)
90-300 nucleotid
Ket hp vi protein tao thanh snRNP (small nuclear
ribonucleoprotein particles), co vai tro trong qua trnh chnh
sa RNA
CHNG 2
I. C s hoa sinh cua qua trnh sao chep DNA
Trc khi phan chia te bao, bo gen phai c sao
chep theo nguyen tac ban bao ton semiconservation. Qua
trnh nay c cong bo au tien vao nam 1958 do Mathew
Meselson va Franklin Stahl
II. Qua trnh sao chep DNA Prokaryote
- 1 replicon
- 1ori C, 1 ter
Khi au sao chep: xay ra Ori C
DnaA + vung oriC (9bp)
DnaB + DnaC + oriC
DnaB trt thao xoan, DnaA phong thch ra ngoai
- Tong hp moi: Enzyme primase + protein primosome
- Tong hp DNA: DNA polymerase III la 1 holoenzyme, gom toi
thieu 10 tieu n v
-DNA polymerase I co vai tro sua sai DNA (5`-3` exonuclease) va
loai bo phan moi RNA khi sao chep
-DNA polymerase II co vai tro cha c hieu chnh xac
- Noi phan oan DNA: enzyme ligase
- Ca 3 loai DNA polymerase : 3`-5` exonuclease
- Loai I: them 5`-3` exonuclease
*** Toc o sao chep E.coli la 1000 bp/s/cha 3 sao chep. o
chnh xac rat cao, ch 1/10
9
-10
10
bp moi the he .
Mot bo may replisome : 2
ban DNA polymerase III
( dimer), 1 primosome va
cac protein thao xoan
Mach cham tao
loop
** Hoat ong Topoisomerase I
Vai tro thao xoan lien tuc trc sao chep
Kiem soat s sieu xoan cua DNA: Prokaryote,
topoisomerase II (DNA gyrase vai tro trong sao chep)
III. Qua trnh sao chep DNA Eukaryote
Thi iem sao chep: pha S
- Toc o sao chep: cham hn,
c 50 nucleotid/s 1 cha 3
sao chep
- Replicon: Tuy cham nhng
qua trnh xay ra ngan la do co
cau truc replicon
- oan okazaki: t 100-200
nucleotid
Co 5 loai DNA polymerase
- ( khi au sao chep ca mach ti va mach cham)
- ( co vai tro sa sai)
- ( = DNA polymerase III - keo dai mach sau o,
khong dimer, ket hp vi PCNA - proliferating cell nuclear
antigen)
- ( vai tro exonuclease dau 3`-5`)
- (sao chep bo gen ti the)
Tng t SSB la RPA (replication protein A)
Tach moi RNA ra khoi oan okazaki nh enzyme
ribonuclease H
Qua trnh sao chep DNA luc lap ?
**Hoat ong cua
Topoisomerase II:
tach 2 mach a sao chep
** Vung Telomere
- O nguoi: 5`TTAGGG 3` ( 200 copies)
** Van e g xay ra Telomere ?
- PP bnh thuong: dau 5` chuoi moi se khong duoc hoan thanh,
ngan dan sau moi lan sao chep
125 lan sc mat Telomere
Te bao sinh dng bnh thng ch NP 1 so lan
roi dng han.
Ngoai tr 1 so loai tb ac biet
Can Telomerase
** Hoat ong cua Telomerase
La ribonucleoprotein, xuc tac tong hp DNA t khung RNA
Tp RNA (TERC- telomer RNA component) tao khung
AAUCCC ( ngi) e tao oan TTAGGG
Tp protein ( TERT - telomere reverse transcriptase) xuc tac
phan ng
Co mat ( bat t do duy tr c chieu dai Telomere):
Te bao mam phoi, trng thanh ( germline)
Eukaryote n bao
Tb ung th ( 85-90% tb tong hp lng telomerase cao)
hng ieu tr
Telomerase va Telomere: yeu to quyet nh i
song te bao
IV. Sa sai DNA
Cac loai ton thng
Mat purin: mat A hay G do lk gia base purin va ng b
nc thuy giai. ay la ton thng xay ra nhieu nhat.
Kh amin: xay ra khong pho bien tac ong cua nc len
lien ket noi nhom amin vi cystosin se tao thanh uracil. Neu
khong sa th se thanh lk AU trong lan sao chep DNA tiep
theo.
Tac ong tia t ngoai hay tac nhan hoa hoc nh benzopyren
trong khoi thuoc la se gay ton hai DNA. Anh hng cua tia
t ngoai se lam 2 thymine ke can noi lai vi nhau tao thanh
thymine dimer.
C ch sa sai
Cat DNA: co 2 loai la cat base va cat nucleotide
Trnh t sa sai: enzyme nhan biet cho sai, ni ton thng b cat, tong
hp lai nucleotid ung, enzyme noi lai vung mi c chnh sa
Khi Cytosin b thay the bi Uracil, U se c nhan biet va chnh sa bi
U-DNA-glycosidase. Enzyme nay se tao mot khoang trong DNA tai
v tr base lk vi nhom deoxyribose. Enzyme AP endonuclease se nhan
biet khoang trong roi cat nhom ng nh pha lk phosphodiester
Khi DNA ton thng do mat purin th AP endonuclease cung cat nhom
ng ma a mat i base purin.
DNA polymerase I se tong hp nucleotid mi va enzyme ligase noi lai
vi nhau
CHNG 3
I. Cac yeu to quan trong
1. RNA polymerase
2. Huong phien ma: 5` - 3`
- Ma di truyen ( ma protein)
- Mach ma ( coding strand)
3. Lien ket DNA-protein
Moi lien ket ben vng do tng tac k nc, lk hydro, lk ion
II. Qua trnh phien ma va ieu hoa bieu hien gen Prokaryote
1. ac iem cau tao RNA polymerase
- Trong lng phan t 450 kD
- 5 loai polypeptid: , , `, , (sigma)
- E.coli, co nhieu loai ,
70
cho phan ln cac gen,
32
cho gen
soc nhiet,
28
cho gen tao roi flagellin
Holoenzyme
2. C che phien ma E.coli ( tap trung vao phien ma tao
mRNA va rRNA)
- Cau truc
Promotor ( 20-200 bp) co 2 vung consensus sequence
* vung -10 (thng TATATT- Pribnow box)
* vung -35( thng TTGACA)
* Qua trnh khi au
- RNA polymerase trt en vung promotor
- oan ngan DNA gan -10 c thao xoan ( vi s
ho tr cua vung pribnow v giau AT nen ch co 2lk
hydro), tao thanh phc hp m.
- Bat au phien ma v tr +1, khi c 10
ribonucleotid th se b phong thch, ket thuc qua
trnh khi au ( bat au bang viec lk gia A hay G
len RNA polymerase, sau o ribonucleotid khac se
en e hnh thanh lk phosphodiester)
- Khi yeu to tach khoi th ai lc gia RNA polymerase
va promotor b giam, qua trnh keo dai bat au
- Core enzyme tr thanh phc hp phien ma, theo hng
5`-3`, DNA pha truoc transcription bubble (TB) bat
au thao xoan dan. Bong phien ma la vung ma 2 mach
DNA thao xoan tam thi, co ket hp gia DNA va RNA
- S thao xoan cua RNA polymerase lam cho vung trc
TB la xoan dng, con sau TB la xoan am. V vay phai
can co topoisomerase
- Tieu n v gan len cac ribonucleotid, vai tro xuc tac
tao lk phosphodiester khi trt tren khuon DNA
- Tieu n v `giup enzyme gan chat len khuon DNA
- Hai tieu n v giup to hp enzyme gan len
promotor giai oan au
* Qua trnh keo dai
* Qua trnh ket thuc
-RNA polymerase se ngng
phien ma khi gap trnh t ket
thuc ( la trnh t co 2 vung giau
G, C, cach nhau mot oan
khong 10 bp), theo sau la mot
oan giau A
-Khi oan nay c phien ma,
th RNA se tao thanh cau truc
kep toc gia 2 vung nay vi
nhau
-Cau truc nay lam cho phc hp
RNA polymerase cham va dng
han
-ong thi, lk gia A va U tren RNA rat long leo, qua
yeu e duy tr TB, nen ca RNA se phong thch ra ngoai,
ket thuc phien ma, cau truc xoan kep DNA hnh thanh tr
lai
Hnh thc ket thuc khong phu thuoc vao yeu to
rho.
-Vai gen E.coli co vung ket thuc khac, c nhan biet
bang protein rho, lam giai phong phc hp RNA-DNA (
la enzyme xuc tac thao xoan gia RNA va DNA, can co
ATP ho tr), no gan len RNA, khong phai RNA
polymerase
Hnh thc ket thuc phu thuoc yeu to rho.
-mRNA la polycistronic
- Chang han operon cha nhieu gen rRNA
polycistronic dai, c metyl hoa
Rnase cat
rRNA trng thanh 16S, 23S, 5S, mot vai tRNA
3. C che ieu hoa bieu hien gen
a. Operon cam ng Lactose
-Vung kiem soat gom promotor, CAP site, operator
- Gom 3 gen cau truc Z ( -galactosidase - xuc tac thuy giai
lactose tao ng n la galactose va glucose), Y(lactose
permease - la chat mang chuyen lactose vao trong tb), A
(thiogalactoside transacetylase)
** Gen i repressor
gan len operator
** Allolactose + repressor
tach repressor ra
operator
khi ong phien
ma
- Operon lac con c kiem soat bi protein CAP
-( catabolite activator protein)
-Glucose la nguon carbon a chuong oi vi E.coli
-Cac enzyme cua operon lac ch c cam ng phien ma vi
toc o nhanh khi khong con glucose. C che nay c ieu
hoa bi 1 phan t truyen tin noi bao goi la cAMP (cyclic
adenosine monophosphat)
-Khi nong o glucose giam, nong o cAMP tang
cAMP ket hp CAP, phc hp nay gan len vung
CAP site
qua trnh phien ma xay ra cang hieu qua
ay la c che ieu hoa dng do phc hp CAP-
cAMP
b. Operon km ham Tryptophan
Gen Tryp R giai ma cho protein km ham dang bat
hoat gl aporepressor. Khi tryp gan len pro nay th mi
tr thanh dang hoat ong gl corepressor
Phc hp nay gan len vung operator cua operon, ngan
can khong cho RNA polymerase gan len vung promotor
khi ong phien ma
Khi lng tryptophan trong te bao giam, phc hp tren
khong c hnh thanh nen se phien ma se c xuc
tien, ket qua tao ra cac enzyme tong hp tryptophan
ay la dang ieu hoa feedback am
III. Qua trnh phien ma va ieu hoa bieu hien gen
Eukaryote
1. ac iem va cau tao RNA polymerase
- Co 3 loai RNA polymerase:
RNA polymerase I hach nhan ( rRNA)
RNA polymerase II tao mRNA va snRNA
RNA polymerase III tao tRNA va 5S rRNA
- Co s ket hp vi nhieu yeu to khac nh TBP, TFIIF, . . .
2.C che phien ma ( chu yeu tao mRNA) va qua trnh
chnh sa sau phien ma
** Promotor
Ln va phc tap
Promotor cua RNA polymerase II co trnh t
consensus, gl TATA box ( -25 en -30)
Co nhng gen khong co TATA box, nhng gen nay ma
hoa cho cac protein can thiet moi te bao gl
housekeeping gen. Promotor cua nhng gen nay gl GC
box
Hoat ong cua nhieu promotor con b anh hng bi
trnh t enhancer - khi c mot protein en ket hp
(transcription factor)
** Che bien sau phien ma
- Gan 5`cap
-Gan uoi poly-A
-Splicing
- Cat chon loc
3. C che ieu hoa bieu hien gen
Mot trong nhng yeu to ieu hoa bieu hien gen la qt methy
hoa DNA ( o cytosin o vung 5`-CG-3` lam ngung phien ma)
va acetyl hoa histon ( gan acetyl va chuoi nhanh cua lysin
histon H3, H4 lam ai lc gia histon va DNA giam, v vay
kch hoat phien ma)
Euchromatin la chromatin t nen, thay ky interphase, la
vung se c phien ma. Tuy nhien, 1 so tb th 1 phan
euchromatin se nen chat hn ( khong bang
heterochromatin), ay cung la 1 c che e ieu tiet bieu
hien 1 so gen nhat nh
a. ieu hoa bieu hien gen c xng
b. ieu hoa bi glucocorticoid
- Hai phc hp thu quan gan len DNA nh 1 dimer. La do
enhancer HRE la palindrome ( mach tren, di co trnh t nh
nhau khi doc tu 5`- AGAACA -3`) - Moi oan 1 mach gl
motif, 2 motif cach nhau khong 3bp. Moi phc hp thu quan
se en gan 1 motif
Nhng chat hoa hoc tiet ra t 1 te bao, lam thay oi
cach bieu hien cac tb khac th gl transmitter - chat
truyen tin ( trong o co glucocorticoid). Cac te bao
sv a bao khi ong hay dng phien ma khi co dau
hieu cua cac chat hoa hoc ngoai bao.
Khong nh hormon steroid, co nhng chat truyen tin
khong dc c qua mang te bao, v vay chung phai
kch hoat he thong truyen thong tin noi bao
(intracellular messenger system) e chuyen chung t
mang vao nhan.
CHNG 4
I. Ma di truyen
- V co 4 loai nucleotid nen se co 4
3
= 64 codon, tao
thanh bo ma di truyen
- Ma di truyen tren mRNA se quy nh trnh t aa tren
mach polypeptid qua qua trnh dch ma. Codon do 4
loai A, U, G, C
** Viec xac nh bo 3 ma hoa cho 1 aa c phat hien vao nam
1961
Marshall Nirenberg va Heinrich Mathew thc hien mot loat
cac th nghiem s dung he thong tong hp protein nhan tao
cha dch chiet E.coli a b pha v tb
Cac polymere RNA tong hp a biet trnh t va 20 aa, ATP,
GTP c bo sung vao
Khi RNA la poly U : san pham la polyphenylalanin (UUU)
Tng t poly A, sf la polylysine ( AAA). Khung la C, san
pham la polyproteinlin (CCC)
Mach khung gom CU lap i lap lai : leucin (CUC)-
serin(UCU)
Trong 64 codon, co 61 codon ma hoa aa, 3 codon vai tro ket
thuc dch ma ( UAA, UAG, UGA) . Trong 61 codon, co 1
codon (AUG) va quy nh methionin, va co vai tro codon
khi ong dch ma
** ac tnh cua ma di truyen:
Suy bien nhng rat ro rang
Methionin va tryptophan la 2 aa duy nhat c
tong hp t 1 codon
Con 18 aa con lai c quy nh bi 2,3,4,6
codon. V vay codon co tnh suy bien
Khong co codon nao giai ma cho hn 1 aa, ieu
nay the hien tnh chat ro rang cua codon
Khong chong lap, khong co cham ket thuc
Trnh t codon tren mRNA c oc bi ribosome lien tuc
t codon khi au en codon ket thuc
Mot bo gom cac codon lien tiep tren mRNA c goi la
khung oc reading frame
Co the xay ra ot bien lech khung, ot bien vo ngha, ot
bien co ngha (missen mutation)
Trnh t th 2 hau nh luon ong nhat, on nh nhat oi vi
cung 1 aa. Aa tch ien, co chuoi ben a nc (v tr
2=purin, A hay G).
V du v tr th 6 globin trong phan t hemoglobin,
khi b ot bien oi GAG sang GUG trem mRNA (chuyen t
glutamat sang valin), lam thay oi toan bo ien tch tong the
mach chuoi cua aa.
Hu qu
Pho quat
Codon co tnh pho quat cho tat ca sinh vat, t E.coli cho en
con ngi
Ngoai le: UGA giai ma tryptophan (ti the); UAA, UAG
giai ma glutamine
(1 so ong vat n bao)
II. Cac yeu to quan trong
a. Ribosome
- Vai tro: ni tong hp protein
- Ribosome co vung gan vao khung mRNA, va vung cho 2
tRNA tch ien. tRNA mang aa se en v tr A, con tRNA gan
vao chuoi polypeptid ang c hnh thanh gan v tr P
Ribosome Prokaryote va Eukaryote: khac kch
thc va thanh phan nhng cung chc nang va hnh
dang ( 2 tieu n v khac nhau, da tren S - n v
lang Svedberg)
Vd: E.coli 70S = 30S ( 16S rRNA + 21 polypeptid)
+ 50S ( 5S rRNA + 23S rRNA + 34 polypeptid)
Eukaryote 80S = 60S (5S, 5.8S, 28S rRNA + 49
polypeptid) + 40S ( 18S rRNA + 30 polypeptid)
b. Tng tac gia codon - anticodon
tRNA la mau chuyen tiep ( adaptor) e dch ma
thong tin di truyen
Moi tRNA mang 1 aa chuyen biet o dau 3` va
anticodon
Nam 1966, Crick a a ra gia thuyet Wobble
giai thch v sao ch co t tRNA can e dch ma 61
codon, da vao cac c s sau:
Thng aa thay oi bo 3 ma hoa v tr th 3
Moi lien ket gia 2 nucleotid au gia codon va
anticodon xay ra nh ly thuyet A=U, G=C
Tng tac v tr 3 khong nh chuan hoa. V vay ch
can co 1 tRNA cho 2 codon khac nhau
Ngoai ra, tRNA anticodon v tr 3 ( vung wobble) co
nucleotid khac goi la nucleoside inosine (I)- co base la
purin hypoxanthin, co the gan vi U, C, A codon
Co toi thieu 31 tRNA khac nhau e dch ma 61
codon thanh 20 loai aa. Tnh them tRNA khi au
se la 32 loai tRNA
Bo gen ty the ( 24 tRNA) va luc lap (30tRNA)
Eukaryote th giai ma tRNA t hn bo gen trong nhan
c. Qua trnh gan acid amin vao tRNA
tng ng
- Thc hien bi enzyme aminoacyl tRNA
synthase, enzyme nay thc hien 2 phan
ung de gan aa vao dau 3` cua tRNA tuong
ng:
Aa c noi vi AMP qua nhom
carboxyl, tao phc hp
aminoacyl-AMP bang cach thuy
giai phong thch pyrophosphat
Chuyen aa t AMP sang tRNA, tao lk ester gia
nhom carboxyl va ca 2` va 3`-OH cua nhom ng
A trong trnh t CCA cuoi moi tRNA e tao
thanh phc hp aminoacyl-tRNA. Nhung ch co 3`-
aminoacyl ester mi c dung trong dch ma.
Bc nay c xem nh bc hoat hoa aa v nang
lng t lien ket phosphodiester nay se c dung
e tao lien ket peptid sau o
P: Acid amin + ATP + tRNA
Aminoacyl-tRNA + AMP + Pi
-tRNA c noi vi aa tng ng: tRNA tch ien
( charged tRNA)
- Co t nhat 20 ezyme aminoacyl tRNA synthase,
moi enzyme thc hien cho moi aa vi tRNA
tng ng
- Enzyme nay co kha nang nhan biet chnh xac
cau truc cua tRNA tng ng e dch ma. V du
gia valin va leucin vi aminoacyl-tRNA tng
ng. tRNA co kha nang chnh sa aa sai do vung
proof reading
II. Sinh tong hp protein Prokaryote
- Toc o nhanh, moi ribosome dch ma khoang
15-20aa/s
- Lien ket peptid c hnh thanh tai 1 vung
chuyen biet tieu n v ln ribosome
Hnh thanh phc hp khi ong
gom ribosome, mRNA, tRNA
IF1, IF3 + tieu n v nho 30S.
IF1 gan len vung A-site, chot vung
nay lai trong suot qua trnh khi
ong toan bo se gan len
mRNA
Trnh t e nh v mRNA gan
vao ribosom 5`-
GGAGG-3` nam upstream 5` cach
codon AUG t 8-13 nucleotid
(Shine-Dalgarno )
Shine Dalgarno lk bo sung voi 3`-
CCUCC-5` o tieu don vi nho 30S
(dau 3`- 1 phan t 16S rRNA)
tieu n v nho v tr ung e
dch ma
** Khi au
Sau o IF2 ( GTP) se en phc hp
tieu n v nho, xuc tien viec gan tRNA
khi au vi codon tng ng tren
mRNA. tRNA khi au Prokaryote la
formylmethionin-tRNA (fmet-
tRNAfmet). tRNA nay co anticodon 5`-
CAU-3`. tRNA nay mang aa se gan o
P-site tieu n v 30S
Qua trnh khi au ket thuc khi GTP
IF2 b thuy giai thanh GDP va Pi, lam
tieu n v ln en gan vao, sau o tat
ca IF se c phong thch
Nh vay phc hp 70S a c hnh
thanh, chuan bi dich ma tu dau 5`-3`
tren mRNA
Gom 3 bc: nh v
aminoacyl-tRNA tren vung A-
site, hnh thanh lien ket
peptid, chuyen v. Qua trnh
nay can co cac EF (elongation
factor)
Aminoacyl-tRNA th 2 en
vung A-site, E= peptidyl
transferase xuc tac hnh thanh
lien ket peptid ( phan 23S
rRNA tieu n v ln
ribosome)
Hai acid amin nay se gan
len tRNA vung A-site(
aminoacyl-tRNA gan vi EF-
tu va GTP). Khi gan c vao
A-site, GTP se b thuy giai
Sau o, EF-ts tach GDP ra
khoi EF-tu, khi GTP khac en
se tach EF-tu ra khoi phc
hp
** Keo dai
tRNA ban au P-site
khong con aa, se b phong
thch ra ngoai va co the tai
s dung
Ribosome di chuyen doc
theo mRNA sang phai e
tRNA mang 2 peptid tren di
chuyen qua vung P-site, A-
site se trong e aminoacyl-
tRNA th 3 en tiep tuc. S
di chuyen nay goi la s
chuyen v (translocation),
can co EF-G-GTP
Qua trnh keo dai ket thuc
khi codon ket thuc vao vung
A-site
polysome
Qua trnh phien ma dch ma xay ra song song Prokaryote
( anh chup di knh hien vi ien t)
** Ket thuc
Qua trnh bat au khi codon ket thuc
en vung A-site ( UAA, UAG, UGA)
Khi o thay v vung nay se gan vi
aa-tRNA tng ng th vung nay se b
lap bi RF (release factor) gom RF-1,
RF-2, RF-3. Khi o chuoi polypeptid
se c phong thch khoi ribosome,
con mRNA, tRNA, cac tieu n v
cung c giai phong
RF-1 phong thch polypeptid t
codon UAA, UAG; RF-2 cho codon
UAA, UGA. RF-3 cha biet ro, vai tro
dng nh ho tr 2 loai tren
** Chnh sa sau dch ma
Prokaryote, gom cac thay oi nh:
Cat protein: loai bo formylmethionin, trnh t tn hieu
(thng nam gan aa cuoi cung, co vai tro quyet nh iem
en cua protein)
Methyl hoa
Phophoryl hoa
Noi vi mot so nhom chc nh: lipoprotein, glycoprotein.
** C che kiem soat dch ma
Prokaryote, cc kiem soat dch ma xay ra mc o
phien ma
Chnh bien the cua trnh t Shine-Dalgarno giup lam
thay oi toc o dch ma cac gen khac nhau ( vd san
pham th 3 trong operon lac ch c tao thanh khoang
1/5 so vi san pham th 1)
IV. Sinh tong hp protein Eukaryote
1. Mot so iem khac biet quan trong
Giai oan keo dai va ket thuc khong khac biet
Giai oan khi au phc tap hn. Aa khi au la
methionin, vi tRNA met tng ng
Sau dch ma, methionin thng b loai bo
mRNA khong co trnh t Shine-Dalgarno e
ribosome gan vao
Do mRNA o dau 5` co gan cap 7-methylguanosine
nen se co 1 so protein gan vao tieu n v nho 40S
e gan len cap nay. Khi gan vao tieu n v nho se
trt doc mRNA e tm codon AUG
Tat ca mRNA Eukaryote eu co trnh t tng t
5`-CCACC-3` pha truoc o ke ben codon AUG (
Kozak), giup ribosome biet a en codon khi au.
Qua trnh nay can toi thieu 9 protein ho tr
2. Qua trnh che bien protein sau dch ma
Thuy giai protein: la thuy giai cat lien ket peptid nh
protease. La qua trnh thng xay ra te bao
Eukaryote.
cat bo methionin au N hay loai bo trnh
t tn hieu
chuyen protein tien chat bat hoat ( pro-
protein) thanh dang hoat ong.
ng hoa li noi chat: a so protein Eukaryote
eu se b ng hoa, la qua trnh gan nhom saccharid
vao protein, la qua trnh chnh sa ong dch ma hay
sau dch ma Eukaryote
Xay ra protein tiet, protein bao quan . . .
Phosphoryl hoa: giup tng tac protein-protein
Hydroxyl hoa: gan -OH
Metyl hoa: gan -CH
3
do methyltransferase xuc tac
Lien ket lipid: Viec noi hoa tr vi nhom lipid vi protein
giup tang kha nang gan vao mang te bao cung nh tng tac
protein-protein
Tao lien ket disulfid: lien ket disulfid thng ch thay cac
protein tiet ( insulin) va 1 so protein mang). Lien ket thng
c tao thanh moi trng khong kh
Cat ghep protein: xay ra sau dch ma. Ket qua tao ra oan
intein khong hoat ong va extein hoat ong
3. nh hng protein
Trnh t tn hieu ( signal sequence):
V tr: au N
au C
gia mach polypeptid
** V du: S van
chuyen protein
qua RER:
-Can SRP
( signal recognition
particle) khi oan
polypeptid u dai,
c 70 aa
Qua
trnh nay c
goi la chuyen
v ong dch
ma.
oi vi trng hp chuyen v sau dch ma
polypeptid c chuyen qua mang la nh
protein hsp70 co gan ATP.
Sau o, trnh t tn hieu se c cat bi
enzyme signal peptidase.
4. Gap khuc protein
Chaperon: Hsp70
Chaperonin
S roi loan gap khuc protein la nguyen
nhan cua 1 so benh ngi nh
Alzheimer (mat tr nh), Creutzfeld-
Jacob (neurodegenerative disease) . . .
5. C che kiem soat dch ma Eukaryote
mRNA di chuyen ra ngoai: phien ma va dch ma tach
biet, giup co thi gian ieu hoa bieu hien gen. S di
chuyen qua lo mang nhan la qua trnh can nang lng,
can co mat cua 5` cap va 3` poly A- tail
Tnh on nh cua mRNA
Kiem soat dch ma am: qua trnh dch ma oi khi b
dng lai do co 1 protein km ham gan len trnh t gan
dau 5`. V du protein ferritin, duoc tong hop khi nong
o Fe te bao len cao
Phosphoryl hoa nhan to khi au eIF
Trt khung dch ma: thng gap te bao b nhiem
retrovirus, ket qua se tao ra hn 1 chuoi polypeptid t
mRNA
V.Anh hng cua thuoc khang sinh len qua trnh sinh
tong hp protein
Chloramphenicol: c che hoat ong peptidyl
transferase
Tetracyclin: c che gan aminoacyl-tRNA len A-site
Streptomycin: c che hnh thanh phc hp 70S khi au
do khong cho tRNAfmet gan vao P-site
Puromycin: lam phong thch sm chuoi polypeptid, tac
ong Prokaryote va Eukaryote.
CHNG 6
I. Phng thc van chuyen protein noi bao
Co 3 cach:
1. Van chuyen co cong kiem soat (gated transport):
- Protein gap khuc en cau truc cuoi cung
- Di chuyen en iem ch
Vd: S van chuyen Protein vao nhan
2. Chuyen v xuyen mang ( transmembrane translocation):
- Protein khong gap khuc hay gap khuc 1 phan
- Di chuyen en iem ch
Vd: S van chuyen Protein en Peroxisome, luc lap,
ti the
3. Bong van chuyen( vesicle trafficking):
- Protein c goi trong cac bong ( vesicle)
- Di chuyen en iem ch
Vd: s van chuyen Protein
+ T li noi chat nham en Golgi (
mat cis)
+ T Golgi xuat bao, en lysozome
II. Van chuyen t nhan
va en nhan
* Cau tao lo mang nhan
* Kiem soat van chuyen ra vao nhan:
nh he enzyme GTPase va chu ky GDP/GTP cua no
Can effector protein : - GAP thuc ay GTPase thuy giai GTP
- GEF trao oi GDP - GTP
Van chuyen ra vao nhan nh GTPase co ten goi la
RAN
- GEF va RAN-GTP: co trong nhan
- GAP va RAN-GDP: co trong te bao chat
** Con ng van chuyen protein vao nhan:
** Con ng van chuyen protein ra ngoai nhan:
III. Van chuyen xuyen mang
1. Van chuyen protein en ti the
- Protein ti the co trnh t tn hieu co the au N ( b
cat) hay gia ( se khong b cat nh Cyt c, a. . . )
40 40
70
22
20
hsp70
23 17
44
hsp70
MPP
TOM
TIM
Mang ngoai
Mang trong
+ Cac protein giup chuyen v mang ngoai: TOM ( translocon of
the outer mitochondrial membrane)
+ Cac protein chuyen v mang trong: TIM
- Cac benh lien quan en roi loan protein ti the: ot
qu, Parkinson, thnh giac kem, tieu ng. . . ( do roi
loan ca gen ti the va gen bo nhan)
2. Van chuyen protein en luc lap
- Trnh t tn hieu au N ( 20 -120 trnh t)
- V du: mo hnh van chuyen protein vao stroma luc
lap
75 75
160
34
hsp70
20 17
110
hsp70
SPP
TOC
TIC
Mang ngoai
Mang trong
hsp70
+ Phc hp van chuyen mang ngoai: TOC ( translocon of the outer
membrane of chloroplasts)
+ Phc hp van chuyen mang trong: TIC
3. Van chuyen Protein en Peroxisome
- Peroxisome co 1 lp mang oi phospholipid - la 1
ngoai le
- Trnh t tn hieu ( au C hay N) se gan vi 1 thu
quan te bao chat
chuyen vao trong peroxisome
IV. S van chuyen qua cac bong gia cac khoang
noi bao
- Exocytotic pathway: t li noi chat, en the
Golgi, en mang plasma, en bao quan
- Endocytotic pathway: t mang plasma en
lysosome ay la con ng ma cac ai phan t
c hap thu va che bien.
1. Qua trnh tach mang va hoa mang
** Bong do lp ao coatomer
ay la c che xay ra trong s van chuyen gia li
noi chat va the Golgi, gia cac chong Golgi rieng re, trong
viec nay choi cac bong tiet t mat trans cua Golgi.
- Lp ao nay gom 7 protein khac nhau ket hp thanh
1 phc hp. Mo hnh hnh thanh lp ao nay Golgi
nh sau:
Ao Coatomer
- Can co GTPase la ARF ( co GEF mang cho e trao oi GTP/GDP)
- ARF-GTP gan len mang cho
- Protein Coatomer en gan len phc hp
- Lam mang tao choi
- Khi en mang ch: GAP mang ch thuy giai GTP, giai phong ARF
va Coatomer
** Bong do ao clathrin
- La phng thc van chuyen Protein mang mannose - 6-
phosphat t trans-Golgi en Lysosome
- Protein thu quan( xen mang) + protein van chuyen ( mannose -
6- phosphat)
- Protein trung gian ( adaptor) en gan vao
- Clathrin en gan len phc hp, lam mang nay choi ( nhng khong
tach c)
Can GTPase la Dynamin ( co gan GTP). S thuy giai GTP tao
lc giup choi tach ra ngoai. Khi o clathrin phong thch ra khoi phc
hp, bong chuan b hoa vi mang ch
++ Qua trnh hoa mang
Can protein SNARES tren ca mang cho lan mang ch
Can GTPase-GTP ( GTPase thuoc ho RAB)
2. Trans-golgi va s tiet protein
+ Protein t Golgi ( mat trans): - tiet ra ngoai ( tiet cau truc
hay tiet ieu hoa)
- en lysosome
+ S khac biet gia:
- Tiet cau truc: Protein c tiet ra khoi te bao lien tuc,
goi la xuat bao
- Tiet ieu hoa: Protein c gi trong cac bong, tch
luy te bao chat
Khi gap tn hieu ( ion Calcium) tiet tch cc ra
ngoai
Van e phat sinh ve kch thc mang tbc, protein xen
mang?
Giai quyet bang c che Nhap bao Endocytosis tao the noi bao
Endosome
a en Lysosome hay Golgi
3. S van chuyen en lysosome
- golgi: gan mannose - 6- phosphat
** Chc nang cua lysosome:
phan giai nhng vat lieu tha cua te bao. e thc hien
vai tro nay, lysosome bac 1 ( primary) se hoa vi mot bong
mang vat lieu tha ( co the en t ben ngoai hay la chat thai
cua cac bao quan ben trong - dang sau con c goi la khong
bao t huy) e tieu hoa chung. Khi nay ta co lysosome bac 2.
Cac vat chat nay co the c phan huy hay van
trong lysosome suot thi gian song cua te bao. Khi o
lysosome nay c goi la the c tru - residual bodies.
CHNG 7
I. Tao dong
1. nh ngha:
Mot dong: tap hp nhieu te bao t 1 te bao goc
ban au
2. Cac giai oan tao dong
a. Tao cDNA ( complemetary DNA)
- Thu nhan, tinh sach mRNA tong so
- Tao cDNA nh enzyme Reverse Transcriptase ( RT
- phien ma ngc)
Dung moi oligo-T
RT
Ribonuclease H
DNA polymerase
DNA ligase
b. a phan t DNA ngoai lai vao trong te bao vi khuan
** Chon loc vector tao dong
V du: Plasmid pBluescript
* Dang au bang: Cat va noi n gian, khong chuyen biet
* Dang au nh: Tnh chuyen biet cao.
V vay phai chen iem nhan biet cua enzyme cat
gii han vao 2 au cua oan DNA ngoai lai
** a plasmid tai to hp vao te bao vi khuan
-X ly CaCl
2
-Bien nap
-Chon dong
mang plasmid
++ Chon dong te bao mang plasmid tai to hp
Dung c chat X-gal va chat cam ng IPTG
Khi co IPTG operon lactose hoat ong co
- galactosidase, lam khuan lac co mau xanh
Khi co DNA ngoai lai chen ung vao plasmid
khuan lac mau trang
Tap hp nhng vi khuan mang plasmid tai to hp tren
goi la Th vien dong hay Th vien cDNA
c. Chon dong cDNA
-Nguyen tac: do DNA hay do protein san pham
** Chuan b th vien cDNA e chay mau do
Trai vi khuan tren a moi trng
anh so
Ap len mang nylon
X ly mang vi chat ly giai te bao
X ly mang tiep vi NaOH( neu dung e do DNA)
** Dung mau do la oligonucleotid ( neu theo nguyen tac do
DNA)
San pham protein biet 1 vai trnh t acid amin
tong hp in-vitro 1 oan oligonuleotid
anh dau phong xa oan oligonucleotid tren (**)
Mang nylon c u vi oligonucleotid phong xa tren
Ra oligonucleotid d tha
at mang len X-ray film, dung ky thuat Phong xa t ghi e
xem ket qua ( om en)
oi chieu lai vi v tr khuan lac tren a cay ban au
Cay ria, gi giong
(**) Phng phap anh dau phong xa mau do
oligonucleotid
- Ngha la phai co gan nhom P
32
- Dung enzyme polynucleotid kinase ( PNK) va c
chat [ - P
32
] ATP
PNK loai bo 5`phosphat cua moi nucleotid, de lai
nhom 5`hydroxyl
Chuyen [ - P
32
] cua ATP sang nhom 5`-hydroxyl
nay
** Dung mau do la khang the ( do protein san pham)
- Dung khang the do tm protein
( nguyen tac nh Western blot)

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