Bước đầu xây dựng quy trình PCR nhằm phát hiện thành phần động vật trong thực phẩm chay dựa trê vùng 16s rDNA ty thể

TAP CHI KHOA HOC TRUONG DAI HOC

MO TP.HCM - SO 4 (37) 2014

3

BUOC DAU xAy DUNG QUY TRINH PCR NHAM PHAT HIEN
THANH PHAN ONG VAT TRONG THUt PHAM CHAY DI A
TREN VUNG 16S rDNA TY THE
Ngiy nhin bai: 10/04/2014
Ngiy nhin lai: 10/06/2014
Ngay duy't ding: 07/07/2014

Lao Dirc Thui)n, Nguyin Thi Than/i Nhan,
Nguyen Thi Thien Throng, Trim Ki'Jn
Vo Phi Phi Nguyen, Phan Thi Tram'
Le Huy'en Afi Thity2

TOM TAT
Hien nay nhu eau su dung thuc pham chay ,igay tang tang,cao
chY trong den "tinh
thulin chay", nghia khong lan bat ki thanh phiin nao.co nguon c ter clang vat. Day mot
dac tinh quan trong trong the biens va san xueit thuc phdm chay.
muc dich kiem tra c6 hoac
khong su hien di en ctia thanh phiin dOng vat trong thtrc phcim chay, chthng toi tien hanh xdy
dung quy trinh phat hien sir hien dien cua thanh phein dOng vat dva tren ki thudt PCR khuech dui
vitng 16S rDNA ty the bang cap moi TP1 va TP2 co .tinh pho.quat va dac hieu voi 16S rDNA
hdu het cac loci Ong vat. Ket qua.cho thdy, buac dau xdy dung thanh tong quy trinh phat hien
DNA ddng vat Min trong thuc pheim chay. Quy ,trinh Avec hinh thanh so. bO nay ctli &roc thin
nghiem tren 11 mdu duac clan nhan la thuc pheim chay thu nhan ter cha va mot s6 tong ty che
bien thuc pheim chay tren thi &yang; Ket qua ghi nhan 4/11 meiu thus philm chay nhiem thanh

phan co nguon goc Ong vat (chiem 36,36%). Qua d6, quy trinh nay co the khong dinh dit ca
khoa hoc de' Olen khai tren mot so hcvng mait low how trong thus te.
Tir khoa: 16S rDNA ty the, thgc phi'rn chay, PCR, Ong vat, th‘rc vat.
ABSTRACT
The demandfbr using vegetarian foods more and more increase nowadays with focusing to
the veganism that mean don't contain any ingredients have origin from animals that is an
important feature in the processing and production of vegetarian foods. For this purpose, we
check whether have or not presence of animal ingredients in vegetarian food. PCR assay that
are specific to detect the presence of ingredients animal were designed basing on 16S rDNA
gene of the mitochondrial DNA genome by primer TP1 and TP2. Oligonucleotide primers that
are universal and specific for 16S rDNA gene in most of animals. As the results, assay was
initially successfully established for detecting animal- origin components in vegetarian foods.
We experimentally tested and detected animal ingredients in 11 samples vegetarian food from
the market and vegetarian food companies, as the results, 4/11 samples (counting for 36.36%)
contain the animal ingredients. In addition, we conducted sequencing and indentified
successfully this ingredient originating from Sus scrota and Gallus gallus. According to this
sequencing result that are completely accordant, we affirm primer-specific amplification in this
study can be apply to experiment on the large number of samples in the reality.
Keywords: mitochondria' 16S rDNA, vegetarian food, PCR, animal, plant.

I
2

Truang Doi hoc Ma TP.HCM.
PGS.TS, Truang Doi hoc Ma TP.HCM. Email:


CONG NGHE

4

2. V4t lieu va phuang phi') nghien

1. Gieri thqu
Thuc pham chay dugc hieu là thkrc pham
kh6ng china cac thAnh phan c6 nguen g6c tir
Ong vatlll. Thuat ngir "an chay" dugc biet den
nhieu a dao Phat va nhieu ton gido kit& dtrai
chay" khac nhauil l. ve xu
nhieu hinh thirc
huang An chay hien nay ngay cang gia tang a
nhieu quec gia tren the giai ding nhu a Viet
Nam. LY do dm skr gia tang nay IA do an chay
IA met phuang thirc trong viec phong
den tim
cac benh tat nguy hiem lien pan den
mach, cao huyet Ap. ung thu,...E21131141. Nam bat
dugc xu the Oat trien, thi tnrang ngay cang
xuat hien nhieu san pham chay vai mh ma da
dang, phong pha nhu: thit ga chay, heo chay,
bo vien chay,... Tuy nhien, "tinh thuan chay"
lai khong dugc dam bao, dac biet là chua c6
met ca quan tharn quyen nAo dam bao trong
thuc pham chay khong IL bat kY met thanh
phan c6 nguen goc dong vat. Do do, met so
cong ty thgc pham chay vai muc dich de dam
bac) "thtrang hieu" phai giri matt thgc pham
(san pham va/hoac nguyen lieu san xuat) sang
nu& ngoAi de kiem dinh. Xuat Oat tir nhu cau
thuc to teen, viec nghien ciru va phat tri6n mot

quy trinh nham kiem dinh sg hien dien co
thAnh phan c6 nguen gee tir dOng vat trong
thkrc pham chay IA can thiet.
phuang phap PCR
Teen the
(polymerase chain reaction) IA phuang phap
thong dung dugc nhi6u tac giA sir dung de
cac thanh phan
khuech.dai trinh DNA dich
co nguOn gec tir Ong vat. Met so cac trinh to
dich dugc sir dung nhu gen Cytochrome b cua
be gen ty the151161, 12S rDNA caa ty the f 71i83,
16S rDNA ty the [9][10]... Trong nghien ciru
nay, 16S rDNA ty the dugc chon lam trinh to
DNA dich cho phan img PCR, ly do cho six lkra
chen nay (1) 16S rDNA c6 tinh phO quat, tinh
bao ton cao trong ding met loci, chang thay
dOi cham theo thoi OW111111121; (2) So lugng
nhieu ban sao 16S rDNA trong to baoi ll i... Do
de, muc dich cifa nghien cuv nay bao gem xay
dkrng quy trinh phat hien thanh phan c6 nguon
gee tir Ong vat trong thut pham chay va buoc
dau quy trinh nay dugc kiem tra tinh thuan
chay tren cac mau dux pham chay thu nhart
tren thuc te.

cum

Veit fieu
Mau chi:mg throng bao gem mL Ong

vat là thit ga va thit heo. Mau chirng am là
mh thkrc vat. Mau thuc to là cac mau thgc
pham chay thu mua ngh nhien tir cac cong ty
san xuat thgc pham chay tir cong ty Au Lac, tir
chg Thu Dau Met NIA sieu thi.
Thiit ke ni'di
Trinh tkr mei xuoi va moi ngtrgc dugc
thiet ke dga teen 16S rDNA ty the ten Ngan
hang Genbank (NCBI: />cifa cac loAi dong vat bao gem ga (KF908854,
AB489247,...), heo (JN714132, KC208030,
KF799977...), bo (KF799979,...) bang cac
phan mem tnrc tuyen nhu Primer3(v.0.4.0)
( Mei sau
khi thiet ke dugc clanh giA cac thong so vat ly
nhu do dAi, nhiet de nong chay, phan tram GC,
nang lugng hinh thanh cac cau true bac hai...
bang phan mem trgc tuyen IDT
( />goAnalyzer/). Deng thoi tinh dac hieu dugc
kiem tra bang chuang trinh BLAST (NCBI:
) v.v.
Tdch chili DNA
DNA dugc tach chit theo phuang phap
Phenol-Chloroformi131[14). Tat ca cac nth dung
de tach DNA deu dugc lan loot rira sach bang
ntrac, con 70° va dung dich PBS (Phosphate
buffer saline). Sau khi rir. a sach, cac mh dugc
cat Nth gia nhuyen thanh met matt dong nhat.
2,0 g mL dich nay huyen phit vai 4 ml dung
dich dong nhat rnAu (NaC1 5M, Tris-HC1 1M,
EDTA 0.5M. ddH2O). Sau khi dong nhat. hat

750 pl dich chuyen vao eng eppendorf mai. be
sung 20 pl SDS 10% va 20 pl proteinase K (1
mg/ml), u qua dem a nhiet de 65°C. Sau de.
250 p.1 NaC1 bao hoa dugc be sung, U matt a
4°C trong 30 phat, ly tam 10.000 vOng/phat
trong 15 phat. Chuyen 500 p.1 dich vao Ong
eppendorf khac va be sung vai 500 pl hen hop
Phenol:chloroform:isoamylalkohol (25:41:1)
dao nhe Ong, ly tam 13.000 vong/phat trong 10
phut (lap,lai 2-3 lan), them 500 pl Chloroform,
dao nhe eng va ly tam 13.000 vong/phitt trong


5

TAP CH[ KHOA HOC TRUONG DAI HOC MO TP HCM - SO 4 (37) 2014

10 phut. Qua trinh ttia dugc thuc hien vai
NH4OAc va Isopropanol a nhiet dO -20°C
trong Yong 2 gia. Sau do, tien hanh ly tam thu
tug va luu gift DNA trong dung dich TE cho
nhang thi nghiem sau nay. Nong d8 DNA
dirge xac dinh thong qua phtrcmg phap do OD
va kiem tra dO tinh sach th6ng qua tri so
A260/A280.

Phan sing PCR
Phan img PCR duce thirc hien vai chu
trinh nhiet sau: 1 chu ky vai nhiet dO 95°C
trong 5 phitt; 35 chu kS7 vai nhiet dO 95°C

trong 30 giay, 67°C trong 1 phtlt, 72°C trong 1
phUt; 1 chu 1(5/ 72°C trong 5 phut. Thanh phan
phan Ung PCR dugc the hien a Bang 1. San
pham PCR dugc dien di tren gel agarose 1,5%
va giai trinh tu tai ding ty Nam Khoa.

Bing 1. Thanh phAn phin tin PCR
Lung

Thanh plan
Master mix (2X)

7,5 ul

MOi xuoi (1012M)

0,5 ul

MOi ngtrgc (101AM)

0,5 .tl

DNA mach khuort

1,0 pi

ddH2O

5,5 gl

Tiing the tich

15,0 p1

3. Ket qui NI thin luOn
dank giti ctic

tinh cut: in'di

Trinh tu mOi de khuech dai 16S rRNA ty
the có trinh tu: moi xuoi (TP1) 5'CCYAGGGATAACAGCGCAATC-3' va mOi

5'(TP2)
ngugc
TCCGGTCTGAACTCAGATCAC-3' (Trong
do, Y thay the cho C va T). Cac dac tinh vat ly
m8i dupe danh gia bang phan mem IDT the
hien ar Bang 2.

Bing 2. Cic dac tinh vat ly ciia mOi TP1 va TP2
INI4i

Chien dai (bp)

%GC

T., (°C)

(1)

(2)

TP1

21

54,7

56,9

0,81

-11,53

TP2

21

52,4

56,2

-2,00

-9,75

(3)
-4,64

Ghi chzi: T,„: nhiet do ming chay: (I) AG cua

frac hp tot: (hairpin-loop) (kcaLmole-i ); (2) AG caa
con true self-dimer (kcaInzolil); (3) AG clia can ink hetero-dimer (kcaLniole.1).
Dip tren ket qua Bang 2. cac gia tri vat
1), ye chieu dai, %GC, nhiet do nong chay va
six chenh lech, nhiet
nong chay de
deft ,thoa
man cac yeti cau trong thiet ke moil'''. Ve AG
phan Ian deu thoa man bam hcrn -9 kcal.mor l,
ngoai trir rang lixgng hinh thanh cau true tu
bat cap dm TP1, TP2. Tuy nhien, khi tien hanh

kiem tra bang phan mem true tuyen, cau true
tu bat cap xay ra a dau 5' va gia tri nay khong
chenh ,lech qua nhieu so yen yeu cau. Do do.
cap mai nay Van dugc sir dung va kiem tra tinh
dac hieu. Tinh dac hieu dugc tien hanh kiem
tra bang chtrang trinh, BLAST va Annhyb deu
cho thay kha nang bat cap dac hieu tren 16S
rDNA a cac loci dOng vat vai mire dO worn


6

CONG NGHE

d6ng dat 100% (Hinh 1) va khOng bat cap teen
cac trinh ttr I 6S rDNA thdc vat nhtx bap cai,
ngo, rau den, dau ht ..: (Dir lieu khong trinh
bay). Dong thai khi kiern ,tra bang phan, mem

ClustalX, ket qua cho thay cap moi bat cap

chuyen biet d6i vt7i cac trinh tu. 16S rDNA tir
d6ng vat (Dil lieu khong trinh bay). Dda tren
nhiing ket qua thu nhan dtrgc, m6i TP1 va TP2
van throe chon de su dung cho cac thdc
nghiem sau nay.

Hinh 1. (a) Su• bat cap cua TP1 va TP2 len trinh tu. 16S rRNA cua Gallus Gallus (Ma so
truy cap teen Genbank: AP003321). Vi tri Y trong cap moi TP1 tuang thich yeri C trong
trinh tu. bat cap; Ket qua BLAST (b) cap mcii TP1 va (c) TP2 the hien tinh dac hieu
hi

lef4

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1343

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1101 TTIAAAATCA CGACCACCTT ACAACCTTAC ACAGCCCCAC TGGGTCCACC

1151 CACACATAAA CCCCTGGTCG ACATTITTCG GTTGGGGCGA CCTTGGAGAA
1201 AAAAAAATCC TCCAAACCCA CAGACCACAA CTCTTCACTA AGACCAACTC
1251 CTCAAAGTAC CAACAGTAAC CAGACCCAAT ATAATTGAGC AATGGACCAA
1301 GCTACES
IMETCCT CCAAGAGCCC ATATCGACAA
1351 GGAGGTTTAC GACCTCGATG TTGGATCAGG ACAACCTAAT GGIGCAACCG
1401 CTATTAAGGG TTCGITTGIT CAACGATTAA CAGICCTACG TGATCTGAGt
1451 #AGACCGGA GCAATCCAGG TCGGTTTCTA TCTATGAACA CACTCCTCCT
1501 AGTACGAAAG GACCGGAGAA GTGGGGTCAA TACCACTGAG CACACCCCAA

moms

(a)

Desoption

Max Tail Ovary E
Mot AcrAssam
Kos sort caw aloe

O c s hum hamtre i4vi Co 13 fritothoocort earOS otnnt

394 394 100% 023 95% 1F342417

• %s
, reffixosus ernathocvw celcott ceetortt

394 39.4 100% 023 95% KF92s379 t

0 Calls Iustis trnhans ritocnonton conYee cenome

394 39 4 100% 023 95% K192i378 I

O Fos tams ribtborienon ornItIt 2enrit

39 4 39 4 100% 023 95% 0925377 t

• ~ylum:anlamcoonwlen ameet ctmemt

39 4 39 4 100% 0 23 95% 0925375

o

39.4 39 4 100% 023 95%
(54.1x.a ce met ai a ^Noronm DNk ctrfti*tt ewmt
lit1i12:tana ir5N.terhVa Paq,eani relnins 1'1+41,11011w CtrIC401 CtnOmt

394 39 4 100% 023 95% tc1921101

(b)
Dexnption
0 pirareercolus smscilus rutothontnon. con iele cenont

Max Totai

store

Glary E

Sart CON nue

dere Accession

42.1 421 100% 0 023 100% U8793001

Pnati *nu rtvfxhandlon, analelt 0e0rit

42.1 42.1 100% 0.013 100% KC533222 I

0 Jilin0010n tan rli1Cdolinn comolet cen0f1

42.1 42.1 100% 0.023 100% KC513221 I

0 Nuts Mtn rreodtntat owl:Me covit

42.1 42.1 100% 0 023 100% 10723381

4cioeis Schtnchl 1140001119n

orree cem Tie

0 Anis DIAMMOOS Mae S 15S r 105cm/11/14 ceni cartiO secuence rvtocii vita

(c)

42.1 421 100% 0.123 100% KCO20795 1
42.1 42.1 100% 0 023 100% 008159 1

7

TAP CHI KHOA HOC TRUONG DAI HOC MO TP.HCM - So 4 (37) 2014
Xily Ong quy trinh thuc nghiem tach
chiet DNA va khukh dpi trinh tfr (rich
'Ring so mau thuc nghiem la 5 tau bao
g6m 2 matt ga (kSf hieu 1, 2), 2 mau heo (kS,
hieu 3, 4) va mau thuc vat (kS, hieu 5), cac mau

nay dugc tach theo phuang phap
Phenol/chloroform. Ket qua tach chiet dugc
kiem tra bang gia tri OD va tS, so A260/A280
(Bing 3).

Bang 3. Gia tri OD va tri so A260/A280 cfia cac mau tach chiet
mAu

OD260

A260/A2R0

CDNA (µg/ml)

1

0,013

1,66

33,55

2

0,063

1,88

158,8

3

0,047

2,00

117,0

4

0,018

1,51

44,0

5

0,028

1,57

70,5

Gia tri A260/A280 cua cac mau tach deu
co gia tri nam trong Idrang_1,8 den 2,0 (cac
mau 1, 2, 3), mot so mau (mau 4, 5) co gia tri
be hcm 1.8, dieu nay nghia la san pham tach

chiet DNA a cac mau nay nhigm protein. Tuy
nhien, cac mau nay van thuc hien PCR khuech
dai trinh to dich. Ket qua dien di san pham
PCR duck the hinh Hinh 2.

Hinh 2. Fet qua din di san pham PCR
ciia cac min nham khao sat kha nAng bAt cap ciia moi

<-149 by

Ghi chti: LAD: thang DNA 100 bp; (-): gang dal chirng am (lchong. chaw DNA).
Ket qua san pham PCR cho thay 6 cac nay cho thay cap m6i TP1 va TP2 dac hieu vai
matt dOng vat: mau 1, 2, 3, 4 deu cc') mot bang trinh to dich 16S rDNA va khong khuech dai
sang ro voi kich thu6c la 149 bp. Dieu nay cho dugc trinh to dich a cac mau thuc vat tren ca
thay moi TP1, TP2 da khuech dai dugc san 1S, thuyet va thuc nghiem. De khang Binh tinh
pham mix tieu 16S rDNA. Trong khi do, 6 dac hieu nay, mot mau san pham PCR dugc
mau thuc vat (bap cai) va a gieng doi chung tien hanh giai trinh to tai ding ty Nam Khoa
am (-) khong thay xuat hien bang nao. Dieu (mau 1) (Hinh 3).
Hinh 3. Ket qua giai trinh tn. mach xuoi (mOi TP1) cfia san phim PCR mau 1
:A

20

30
is
- • __ . • --mgr.
GTTTAC GACCT GC kT

G, T CO AC. GG

120
110
90
100
80
0
60
50
40
-s••••••ses•a•• - •
ZCAGG 1CACCT ATGGIGCAACC GCT.T 7;,AGGGTTC GT TTGTTCAACG ATTAkCAGT GCTACGTG ATCT GAGTTCAGACCGGR4 T

fi

V,N yw,Arpf ou„


8

CONG NGI4

Dina ten ket qua giai trinh to Hinh 3, ket
qua cho thay cac peak dm san pham deu rat,ro

rang chi trir met doan so nucleotide phan dau.
Dieu nay co the giai thich, tai vi tri doan mei
bat cap, khi giai trinh to tin hieu vung nay
khong re rang. Tut' ,nhien, phan d9c dtrac
(doan
trinh ttr) rat ro rang va hoan toan
dac hieu khi dupc tien hanh kiem tra bang
chuang trinh BLAST. Ket qua BLAST cho
thay, doan trinh to nay ttrcmg (tong rat cao vori
trinh to 16S rDNA ty the dm loci Gallus gallus

(Ma so troy cap: KF981434) vet dO ttrong
deng dat 98%, gia tri E-value gan bang 0
(Ident=98%, E-value=4e-46). Ngoai ra, chang
toi tien hanh kiem tra sir tuang &mg ved cac
trinh to 16S rDNA cua ga thu nhan to Ngan
hang Genbank deu cho thay do arc:mg Tong rat
cao (Hinh 4.). Do do, ket qua trinh to 16S
rDNA nay tuang Tong v6i loci ga (Gallus
gallus) hay not cach khac DNA tach va khuech
dai dung yen mau thit ga.

Hinh 4. Ket qui sip giong cat trinh tkr san phim PCR-TP1
voi cac trinh fir 16S rDNA ciia ga (Gallus gallus)
Iltttettilltft

A8489247.1
KF908854.1
AB489247.1
AY236430.1

Sanpham TP1

A TACAA A
7AC:ACC 7CA 7;'7
CA:',:ACAACC AA
C..AAX.X7A7 -AA
- C.AACX7176CA:,7CC:AC.. - A .1. 7CVACC:•:14
C ACAL. A "'AC ACC7CA 7:4 TAYACALT AA
:MCC:CA AA
CAKA7A/CA=7AC A
A •CA =A
A"C:ACAA,,
7ACACCTII 7c.7:A-CA YACALCC7 AA
"CAICIA716ACk;7CC-AC: A :
:A :ACC
-CACAVACP777CACCTCA . f.:A!"CX,IICAACC7AA ..CAACCX*177MX•77C.,
7AACA:7CCAC .1 A
A A .AX.:24
1170.1ACAA.Y.AX,717Ar..ACC7C..1 , . ,....ATCACZACAACC7M-....:7.CAAC017AT7AAV
.417CJ ...,"."7CAACSA77AACI7CCTAC 7-- A
A "C.A.ACC:a

Nhu vay,,
tren cac ket qua danh gia
1S, thuyet va ket qua, thuc nghiem, chung toi
nhan thay da bulk dau thanh cong trong vi'ec
xay dtmg quy trinh thuc nghiem khuech dai
doan trinh to dich 16S rDNA nham phat hien
thanh phan dOng vat hien dien trong thuc
pham chay.

Kb qua that nghiem tren cac mau

dire d

Sau khi xay dung ducc quy trinh thtrc
nghiem, buerc dau tien hanh thin nghiem tren
11 .mau thu mua clic, ding ty san xuat thuc
pham chay va sieu thi ten dia ban Thu Dau
Met. Ket qua kiem tra duck the hien ten bang
dien di Hinh 5.

Hinh 5. Ket qui kiem tra sty hien dien thanh phAn Ong vat tren cac mau dug nghiem

Ghi cha: (I) (2) (3) (4) (5) la cac mau thu nhan tic cfra hang san xuk thuc pham chay Au Lgc Ian lwat la
dai get xa chay, thit ngc kho chay, swan ram chay, bo kho chay, pate gan ngong chay; (6) (7) (8) la cac
mau thu mua tai chq lan Iwo la heo chien chay, chci ming vit chay, chci ca chay; (12) (13) (14) la cac
mau thu mua tai sieu thi kin Iwo la ga cyc chay, bqt xa va thit gel xe chat'; (9) (11) (15) la chang throng
lan lwat la this heo, thit ga, hon hop tran this ga va thit heo; (11) la doi chcing am, ccii bap; (-) gieng
chitng am khong china DNA.


TAP CHI KHOA HOC TRUONG DAI HOC MO TP.HCM - SO 4 (37) 2014

Dkra tren kEt qua Hinh 5, cac mau thkrc
phdm chay a chg deu xuat hien mot bang a
kich thtrac 149 by (3/3 mau niam); a cac mau
thu nhdn tir sieu thi, có duy nhat I mau (mau
so 12) (1/3 mau nhiEm) xuat hien mot bang
149 by voi (10 sang ttrang dal ma; a cac mau

thu nhdn tix ding, ty san xuat thkrc phdm chay
Au Lac khong thdy xuat hien bang nao ca (0/5
mau nhi6). DO sang khong Tong dEu a cac
mau bi nhiErn dtrgc giai thich la do ham luting
protein (Icing vat nhiEm trong cac mau khac
nhau. ming dO DNA tach &rot khong giong
nhau, dAn dEn sir khuech dai cac bang có do
sang khac nhau. DOi voi nth chimg dtrang
deu hoan toan xuat hien bang vai kich thtrac
149 by sang, mau chi ng am khong xuat hien.
Nhtr vdy, tj, re so mau nhim protein dOng vat
dat 4/11 mau chiem 36.36%, trong do cac mau
cha ty le nhiem 100%., sieu thi nhie'm
33.33% va cong ty san xuat nhrEm 0%. DE
khang dinh han ve ket qua, mot mau dai dien
chive tien hanh giai trinh ttr co xuat hien bang
(mau so 7). Ket qua giai trinh tu. &lac kiem tra
bang chtrang trinh BLAST. KEt qua kiem tra

9

BLAST cho thAy, mau san phdm tuong &Ong
vai I6S DNA cua Sus scrolit vai gia tri ttrang
clang cao dat 98% va gia tri E-value gan bang
0, (Ident=98%, E-value=4e-51) va dO arcing
dong cao vai cac trinh tir 165 DNA ctia heo khi
kiem tra bang chtrang trinh ClustalX (Dix lieu
khong trinh bay).
4. Kat ludn
Quy trinh Oat hien thanh phAn có nguon

thudt sinh hoc
goc tir dOng vdt dua tren
phan tir PCR tren gen nwc tieu 165 DNA da
buck Tau xdy dung thanh cong vai dO dac hieu
cao. 'long that quy trinh dugc ap duns de.
kiem tra thir nghiem tren cac mau thkrc to thu
nhdn tren dia ban ThU D'au,MOt, Binh Duang.
Ket qua btrac &du cho thdy ty le nhiem dat
36,36% (4/11 mau nhiEm). DE img dung quy
trinh nay rOng Cal trong thgc te. trong, cac
nghien cat sap tai. chung,toi tie') tuc toi tru
he quy trinh tach chief doi voi nhieu loai, mau
(thkrc pham khac nhau) ding nhu• thiet ke
chimg not nham khang dinh tinh tin cay cua
quy trinh.

TAI LIEU THAM KHAO
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10

CONG NGHE

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