5.

5.

450
- ()
--2 (-2)

(screening) DNA

50 :

190

5.

.
50
,
.

1950, 6-
(G6PD) . G6PD
, NADPH. NADPH
shock. ,
G6PD .
, shock,
.
.
G6PD,
. G6PD
, 400 135
.

,

.
,
, >1%.

,
(SNP: single nucleotide polymorphism).
1 1.330 .
SNP.
SNP ,

191
( )
>10 SNP, >1%. > 1
.

.

.
,
.
CYP3A4.
, - ,
Ca2+,
CYP3A4, , . ,

.
,
.

.
CYP3A4, .

.
,
.
microarrays (DNA chips) ,
,
.

. PCR (Polymerase Chain
Reaction) RFLP (Restriction Fragment Length Polymorphism),
SSCP (Single-Stranded Conformation Polymorphism) TGGE/DGGE
(Temperature or Denaturating Gradient Gel Electrophoresis). ,
.
DNA chips ( DNA microarrays gene chips)
. Northern Southern blots
.
,
mRNA cDNA .
chips /
, SNPs (Singlenucleotide Polymorphism), .
, cDNA microarrays (
Brown Stanford, Shena et al, 1995) arrays
( GeneChip
Affymetrix, Inc, Santa Clara, CA, USA, Agilent Technologies, Palo Alto, CA,
USA):

192
cDNA microarrays cDNA ,
PCR, (chip)
30mm x 15mm. 50-150m, chip
30.000 , .
(
mRNA) . (.
Alzheimer )
mRNA (600ng mRNA chip 10.000 ),
DNA (complementary DNA - cDNA),
. , cDNA
(. Alzheimer,
). ( Alzheimer) chip,
laser .
,
: , :
, : ( 5.1).

mRNA,

cDNA

cDNA

/database

Scanning

5.1 cDNA microarray.

1. RNA ( ). 2. RNA
3.
cDNA. 4. cDNA chip. 5. laser
. D. Geschwind, DNA microarrays:

translation of the genome from laboratory to clinic, The Lancet, 2, 275-282, 2003.

193
.
chips - -
.
1. chips ( 25
) in situ
(GeneChip), chips
ink-jet (Agilent Technologies).
, Stephen Fodor 1994,

5 ,
. ,
, ()
, ,
,
. chip (),
.
,
. 4x25
425 25.

5.2 chip .
5
( ),
(
() . (),
.
(G, A, C)
. 1,6 cm2 65.000 - 400.000
. Biochemistry, Voet and Voet, 2004.

194
chips 65.000 - 400.000 .
, chip .
(

), chip 1,6cm2 9.000
. 20 (
25), ,
.
, ,
,
.

5.3 () chip
9.000 . 20 25-.
25 ,
, .
40 ( 9.000 360.000 ).
() 25, cRNA
(perfect match), (mismatch).
20 , 20 ,
.
Lipshutz R.J., Fodor S.A., Gingeras T.R, and Lockhart D.J., High density synthetic

oligonucleotide arrays, Nature Genetics, 21: 20-24 (1999).

/
chip , . 350.000
40.000 bp ,
DNA
. (25 17),

( 1 , 2 , 3 G, 4 C).
cRNA

. chips
, 450,
.

195

5.4 . chip /
1 2 . chip (0.8 x 0.8 cm)
HIV (HIV PR) (HIV RT). 1515
. .
.

. () .

Lipshutz R.J., Fodor S.A., Gingeras T.R, and Lockhart D.J., High density synthetic oligonucleotide
arrays, Nature Genetics, 21: 20-24 (1999).

5.5
chips. () Chip

Affymetrix () chip
cDNA.

cRNA
cDNA.




RNA DNA.


- ( chip
Affymetrix =
, =
). Gerhold D.,

Rushmore T., and Caskey C.T.,

DNA chips: promising toys
have become powerful tools,
TiBS 24: 168-175 (1999).

196
.
chip .
3-5g mRNA,
cDNA.
cDNA, 7 SP6 mRNA
cDNA cRNA.
cRNA . cRNA-
DNA chip
.
/
DNA cDA, PCR.
DNA
.

chips
( 175.000$)
chips,
,
/ ,
/ , .
, chips Affymetrix
450 (
5.6)
( 5.7).
5.6 chip
Affymetrix

(Cyp)
. Gerhold D., Rushmore

T., and Caskey C.T., DNA chips:

promising toys have become powerful
tools, TiBS 24: 168-175 (1999).

5.7
chip Affymetrix


:
(ApoE), PSA (prostate specific
antigen), Trpmy1E (trpopomyosin
1E) . Gerhold D., et al., DNA

chips: promising toys have become

powerful tools, TiBS 24: 168-175
(1999).

197

.

.
. ,
, ,
, .
,
,

. ,
.

,
, .
CYP2D6
. CYP2D6 40
. tests
, test , ,
.
.
, 4--
.
0.2 - 12.6 .

.

(over-dose).
5.8.

R=

4--

(MR: metabolic ratio)

5.8 CYP2D6 .

RFLP-PCR.
70 .

198
CYP2D6*4 1934,
3 4.
181 457 .

CYP2D6*2.
.

.

.
450, ,
. ( 5.9). ,
,
.

5.9 .
450 (CYP), (DPYD)
, - (UGTs), -S (GST), -- (s) -
().

450
450
.
450 - ,
.
-. -
, . ,
450 .

199
5.1 4

(CYP) (CYP1-CYP4),
( CYP2A, CYP2B, CYP2C, CYP2D, CYP2E, CYP2F)
1-20 (. CYP2C9).
(CYP2C19*3).
: CYP3A4, CYP2D6, CYP2C9, CYP2C19 CYP2A6.
, 5.1.
5.2 ,
.
, CYP2D6, . (induction)
- .
.
internet.
5.2 .

CYP2C9

NSAID (non-Steroidal anti-inflammatory drugs)

CYP2C19

CYP2D6

200
- ()


, Crohn. , 6 (6-) .
.
6-. -
(HGPRT) 6- 6-G (6- )
.
6- . ,
, 6- 6-.
. 6.
8 .
*2, *3 *3C. 0,3 %
( ).
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. , 6-
(6-GN)
.
:
: --
:
o (Crohn).

6-:

TPMT: --
6-:

HGPRT: -

8 :
*2
*3
*3C
0,3%

6-TGN: 6- :

.
: 6-
6- HPLC.
.

201
--2 (-2)
-.
1952.
1953 ,
.

-.
.
( 5.10).

5.10
, .
, . 267
,
: ( )
( ). Weinshilboum R, and Wang

L., Pharmacogenomics: Bench to bedside, Nature Reviews Drug Discovery 3, 739-748 (2004).

, . 1991,
cDNA -2, ,
NAT2*5 2*6. 90%
. 2003, 14
-2, 36
.
40:60
, 85:15 95:5 . .

.

.

202
NAT-2 ,
AFMU/1-,
. 45,3% 54,6% .
55,8% /, 39,2% / 4.9%
/. 5,7% ( 5.11).

5.11 -2
(AFMU/1-), .
Weinshilboum R, and Wang L., Pharmacogenomics: Bench to bedside, Nature Reviews

Drug Discovery 3, 739-748 (2004).

.
10
. ,

203
. ,
(Balant-Gorgia et al, 1990).


,
, .


,
.

.
, SSRIs (specific serotonin reuptake inhibitors:
),
.

17q11.1-q12.


. : , 14
23 , 16 .
mRNA .

SSRIs.
2
9, 10 12 17
.
.
SSRIs.


, .
D2. :
. ,

. ,
,
(agranulocytosis). , D4
D2.
5-2.
D4 ,

48 . D4x,
.
,
4 . ,
- 5-2
.

204

.
.
,

.
50%
, Ser9Gly
D3.

,
.
.
, , ,
.

, .

: , .
,

. , -
, . ,

.

1990, Lee et al -
,
-
.
CYP2D6,
( ).
*2 *3 CYP2C9
.
, ,
(. ) .
. - - -
CYP2C9.

. -
-
.
-
26 (C345T) MDR1C.

205
. ( 24%
) ( ) 4
.
.

5.12 - -

.

.
LQTS (long QT syndrome)
.
- .

5.13 () (r
IKs) LQT (long QT syndrome)
.

206

.
, .

-,
.
, ,
, UGT1A1.
- SN-38,
.
SN-38
.
([]n) UGT1A1,
n.

.
. (
, , .)
.
.
,
. ,

. ,
, . ,

(nucleus
accumbens).
.
: 1/
5 (ADH) , 2/
.
.
-
(, , , .).
.
ADH2 His47 , ALDH2
Lys487 ,
.
.
( ALDH2*2)
.

.
, -

207
DRD2 1 ,
. 1
,
D2.
, ,
,
.
, ,
.
,

.
, . , Batra et al (2000)

DRD2 Noble et al (1994) Comings et al (1996)
.
.
, 3 ,
, .

.
(, , , , ). SNPs
. ,
Gaba-
.

. . 118G ,
10% ,
- .

.

, ,
.

.
30-40%
. in vitro
: 450,
, .

, a priori
. ,
, .

208
Murphy et al (2000) CYP2D6
,
. , a
posteriori .
CYP
. , .
, .
,
.
,
.

35.000 , 43
100 .
. , GPCRs

, , , ,
, .


. ,
(target validation)
.


.
(Plasmodium falciparum),
(Anopheles gambiae) (Homo sapiens)
.
,
.

.

. (-/-) (collection of yeast
deletion strains).

.
,
(6000 ).
(replacement cassette),
TAG (
),

209
. TAG
(barcode), .
ATG-

upTAG

BARCODE

dnTAG

-TAA

5.14 . barcode TAG .

. :
, DNA.
DNA.


.

.
(screening) DNA
, ,
,
DNA, .
.
,
.
,
,
. :
deletion strains.
,

. 96
. ,

. .
130
. 107
.
.
,
deletion strains. ,
.
barcode, .
.

. PCR (
primers ) barcodes
,

210
,
.

, .
, ,
,
. 35%.

5.15

.
(-/-)
,
.
PCR

barcode.

,
,
, .
, (rapamycin sirolimus)
yeast deletion collection. , ,
(downstream) .
.
,
FKBP12. -FKBP12
Tor1p Tor2p
FRAP/RAFT/mTOR,
(IL-2) -, . Tor/FRAP
.

211

5.16 (Sirolimus).
(proliferation) -. Goodman & Gilmans Pharmacological Basis of

Therapeutics, McGraw-Hill, 2001.

,
,
(.. , , ).
Tor .
,
(wortmannin), Talaromyces
wortmanni. -
. PIK (PI kinase),
() 4,5
(PIP2), (
PKC), - (
).
PKC,
-,
.
- ,
, 2.
.

( IP3), ,

212
,
-.

,
DNA. 1000
,
.
2003.

(6000 ) (high-throughput
screening).
.

, ,
,
. , , .

.

. ,

.

.

1- .
, ,
, .

. , -
.
,
1
.
, . 1R
Arg3891AR Gly3891AR.

- .
Arg3891AR Gs
Gly .

1
, Arg3891AR Gly3891AR,

. :

213

(3 ) Arg3891AR
, , Gly3891AR ,

( 5.15).
, Arg3891AR 6 ,
( 5.15).
Arg3891AR .
(3 ) -,
Arg3891AR , Gly3891AR
( 5.15).
(3 ) -
(), 5
Arg3891AR , Gly3891AR (
5.15).
Arg3891AR
- Gly3891AR.
.

5.17 1 (Arg3891AR Gly3891AR):

. . (3 ) Arg3891AR
- , Gly3891AR
. . 6 Arg3891AR . .
3 Arg3891AR
. . 4 Arg3891AR
, ,
Gly3891AR. Ligget S.B., Genetically modified mouse models for pharmacogenomic research,

Nature Reviews Genetics 5, 657-663, 2004.

214
, Arg3891AR
Gly3891AR,
. , -
Arg3891AR , Gly3891AR.
5.18 1
(Arg3891AR Gly3891AR)
-.
Arg3891AR Gly3891AR,

(-): Gly3891AR
,
Arg3891AR .

Ligget S.B., Genetically modified mouse models

for pharmacogenomic research, Nature Reviews
Genetics 5, 657-663, 2004.

50 :

.

.
5.19

PUBMED
(National Library of Medicine).
Vogel
1959,
1998.
5
. Meyer U.S.,

Pharmacogenetics five decades of

therapeutic lessons from genetic diversity,
Nature Reviews Genetics 5, 669-676 (2004).

,
, .
.
:
1) .
2)
.

215
3) ,
.
4) .
5)
.
6) .
7)
.

. /
. , kit

.

-
.
1892, o Sir William Osler
,
.

,

.

, gefitinib,
EGF. , 10%
. ,

. ,
.
: gefitinib,
EGF, .

.

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