心境障碍与5-羟色胺转运体基因的连锁不平衡
维普资讯 http://www.cqvip.com
ISSN 1671—5926 CN 21—1470/R
w .zgfc com kf23385083@sina.com 高树赛,等.心境障碍与 5一羟色胺转运体基因的连锁不平衡
DNA polymerase (1 U), 2×GC buffer I (10 L), and the two
primers(both of 1 0 mmol/L).The reaction was performed in 2 cy—
cles of predenaturat...
维普资讯 http://www.cqvip.com
ISSN 1671—5926 CN 21—1470/R
w .zgfc com kf23385083@sina.com 高树赛,等.心境障碍与 5一羟色胺转运体基因的连锁不平衡
DNA polymerase (1 U), 2×GC buffer I (10 L), and the two
primers(both of 1 0 mmol/L).The reaction was performed in 2 cy—
cles of predenaturation at 94℃ for 3 minutes.94℃ for 30 seconds,
63℃ for 30 seconds.and 72℃ for 1 minute.2 cycles of 94℃ for
30 seconds,62℃ for 30 seconds,72℃ for 1 minute,and 34 cy—
cles of 94 ℃ for 30 seconds. 61℃ for 30 seconds. 72℃ for 1
minute foIlowed by a final extension at 72℃ for 6 minutes.( 5一
HTrvN ITR loci:Th e total reaction system was 20 L containing DNA
template(50 ng genomic DNA),TaKaRa Taq polymerase(1 U),two
primers(both of 10 mmol/L),and lOx PCR buffer(2 L).After
predenaturation at 95 oC for 3 minutes,the reaction was perform ed
in 34 cycles of 95℃ for 1 minute.55℃ for 45 seconds.72℃ for 1
minute with a finaI extension at 72℃ for 10 minutes. ( 5一HTT一
3’UTRG/T Ioci: Th e totaI reacting system of 20 L¨ contained
DNA template(50 ng genomic DNA),Taq polymerase(1 U),the
primers (both of 10 mmol/L), 10xPCR buffer (2 tLL), and 1.5
mmo1/L MgC1’. Atier predenaturation at 95℃ for 3 minutes, the
reaction was perform ed in 35 cycles of 94℃ for 30 seconds.56℃
for 1 minute. and 72℃ for 1 minute with a finaI extension at 72℃
for 6 minutes. MseI endonuclease was added for digestion at 37℃
in water baths overnight. Th e restriction enzyme system in a totaI
volume of 1 0 L consisted of PCR product (2 L), MseI (2 U),
10xbuffer R+f1 IxL1 and adequate amount of ultrapure water.
Gel electrophoresis and genotyping:The PCR product of the 3 loci
fincluding the digestion products of 5一HTT3’UTRG/T Ioci1 under—
went 6% nondenaturing polyacrylamide gel electrophoresis(PAGE)
for separation for 3-4 hours, and stained with silver nitrate. Th e
genotypes of the 3 Ipci were determ ined according to the patterns
of the bands displayed by electrophoresis.
Statistical analysis:TDT was used for linkage disequilibrium analy—
sis. In the core families comprising the patients and their parents,
the probability of transmission of an allele from the parents wi th
norm aI phenotype to their offsprings with mood discrder were eval
uated with the parents as the internal contro1.In this study.extend.
ed transmission/disequilibrium test伍TDrn software was adopted for
statisticaI analysis of the association of each allele of the Ipci and
the haploids with different combinations of the alleles with mood
discrder[61. Th e laboratory experiment and part of the statistical
analysis were completed in the Department of Forensic Medicine,
College of Basic MedicaI Sciences.Sun Yat—sen University.
RESULTS
Quantitative analysis of participants
Altogether 72 families, 247 subjects, were enrolled in this study
including 65 COreS families containing 193 subjects 【patients and
their parents, but 2 families only had a father), 4 sibling families
with 17 subjects(more than 2 siblings were affected),and 3 high—
risk family involving 37 subjects(over 3 members in 3 generations
within two genealogies were affected).Of these subjects,232 in 72
core families were eligible for hereditary correlation analysis and
divided into④ patient group with 90 patients consisting of 5 1
male and 39 female patients aged 15-51 years with a mean of
(24_+7)years,whose onset age varied from 12 to 35 years with an
average of (20_+5) years, the course of illness ranged from 0.5
month to 1 8 years with the median of 4 years.In these patients,56
had bipolar disorder,and 34 had major depression(including sin—
gle—episode depression and recurrent major depression).( Parents
group,including 142 parents consisting of 72 fathers and 70 moth—
ers aged 36—78 years with an average of(5 l+8)years.
Statistical inference
ETDT analysis of 3 polymorphie loci in serotonin transporter gene
(Table 1)
Table 1 shows that in the 72 core families, the empirical P values
171
of the 3 loci did not exhibit statistical significance,suggesting that
there was no linkage disequilibrium between these 3 loci and mood
discrder.
Table l Result of transmission/disequilibrium test of serotonin trans—
porter gene f5一H1Tn allele in 72 mood disorder core families
Transmission or non—transmission is the number of families in which the heterozygous
parents tran s mitted the alleles to their offspring. The P value is the empirical value
after 1 000 corrections with Monte Carlo sireulation; The detected 3 alIdes in 5.HT.
TI|PR loci by PCR were s,L'L+ with length of 484 bp,528 bp,528+bp,respective—
ly;alneligible for ETDT test.
Analysis of haploids with different combination of the alleles at the
loci of 5-HTT gene(Table 21
Table 2 Analysis of haploids with different combination of the alleles at
the loci of serotonin transporter gene (5-HTT) gene in 72 mood disorder
corefamilies
H
⋯
aploids
.
w
.
ith di雎 ren T
ransmissi0n Non.transmission P
aLL eLe cotnblnatlons
5.H 【’rI R.VNTR
S 【2 25 29 O_3O 0.58
S 【O 1O 8 O.22 O.71
L I2 2O 15 O.71 O-39
L 10 6 7 008 0 78
L 12 O 2
5.H‘ 【’rI R一3’UTRG/r
31 25 0.64 0.42
r 17 26 1.84 0.17
L : 22 14 1.78 O.18
I』 r 4 7 O.82 O.36
L C O 2
5 H‘ rrVNTR.3’UTRG/r
1: !G 31 17 4.O8 0.04
1: 1T 14 24 2.63 O.11
1I )p 3 5
1I l1 13 O 17 0 68
5.H‘ 厂T1PH.VNTR.3’U.rRG/r
S 2G 31 22 1.53 O.22
S 2T 12 21 2.46 0 12
S OG 4 1
S 0T 5 9 1 14 0.29
L 2G l9 l0 2.79 0.09
L 21、 1 5
L 0G 2 4
L 0T' 4 4
L l2Ga O 2
Transmission or non.trails mission was
gous parents transmi tt~ the alleles in
Ineligible for multi—allele TDT.
the number of families in which the heterozy—
different combinations to a haploid offspring.a
Table 2 shows that of the 72 core families, haploid formed by the
combination of 5.HTT VNTR and 3’UTRG/T was found connected
with mood disorder =4.08,empirical P value of 0.04,with 1 000
corrections by Monte Carlo simulation),but linkage disequilibrium
could not be observed in the haploids with other loci(尸>0.05).
DISCUSSIoN
Human 5一H I gene locates in the region 0f 17q11.1—12 coding f0r
5一H I protein, which pIavs a kev roIe in serotonin reuptake into
the presynaptic neurons闱.OgiIvie e£ⅡZ examined the reIationship
between poIvmorp sm of VNTR in the sec0nd intr0n of 5一HTT
骶me and mood disorder,and f0r the 6rst time demonstrated the as—
sociation between 5.HTT gene and maior depression.Lesch e£ 【句
aIso studied the insertion/depIetion Ien甜h poIymorphism of a l4一
bp flra ent 1 kb upstream of 5一HTT#rene, and discovered that 5一
HTTLPR w_ds obvious connected with anxious personalitv. In a
case.connl(】lled coITelation studv of mood dis再 er and 5.H I n.P
carried out bv CoUier e£ [7】 based on the data collected f om 3
维普资讯 http://www.cqvip.com
维普资讯 http://www.cqvip.com
本文档为【心境障碍与5-羟色胺转运体基因的连锁不平衡】,请使用软件OFFICE或WPS软件打开。作品中的文字与图均可以修改和编辑,
图片更改请在作品中右键图片并更换,文字修改请直接点击文字进行修改,也可以新增和删除文档中的内容。
[版权声明] 本站所有资料为用户分享产生,若发现您的权利被侵害,请联系客服邮件isharekefu@iask.cn,我们尽快处理。
本作品所展示的图片、画像、字体、音乐的版权可能需版权方额外授权,请谨慎使用。
网站提供的党政主题相关内容(国旗、国徽、党徽..)目的在于配合国家政策宣传,仅限个人学习分享使用,禁止用于任何广告和商用目的。