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M HUY HONG
Ngi hng dn khoa hc: TS. Nguyn V Thanh Thanh Thi Nguyn-2011
4. KT LUN V NGHI
Phn lp v xc nh trnh t gen DREB1 mt s ging u tng c kh nng chu hn khc nhau
CYS
VNS
2.2. Ha cht v thit b 2.3. a im nghin cu 2.4. Phng php nghin cu 2.4.1. Phng php tch chit DNA tng s
1
2 3
2,5
2,5 1
4
5 6 7 8
1
0,25 3 2,5 12,25 25
1 2 3 4 5 6
94 94 51 72 72 4
30 1
2.4.4. Phng php tinh sch sn phm PCR Sn phm c tinh sch (thi gel) theo b Kit AccuPrep PCR Purification (Bioneer-Hn Quc) 2.4.5. To vector ti t hp Bng 2.6. Thnh phn phn ng gn gen vo vector
STT 1 2 3 4 Thnh phn phn ng T4 Ligase Buffer T4 DNA Ligase pTZ57R/T Vector PCR product Th tch (l) 1 1 1 5
Nc ct kh ion
Tng
2
10
1
2
1
0,5
3
4 5
XbaI enzym
DNA plasmid Nc ct kh ion Tng
0,5
3 5 10
KT QU V THO LUN
3.1. Kt qu tch chit DNA tng s
1 1 2 2
Hinh 3.1. DNA tng s ca 2 ging u tng CYS v VNS. Ky hiu: 1: CYS; 2: VNS
Tn ging A260
CYS
VNS
A280
0,170
0,128
A260/A280
1,911
1,813
0,325
0,232
717bp
Hinh 3.2. Kt qu in di sn phm PCR nhn gen DREB1 ca 2 ging u tng CYS v VNS. Ky hiu: M: Marker; 1: CYS; 2: VNS
2800 bp 717 bp
Hinh 3.5. Kt qu in di sn phm ct plasmid ti t hp bng BamHI v XbaI M: Marker 1kb; 1: CYS; 2: VNS
Hnh 3.6. So snh trnh t nucleotide ca gen DREB1 CYS vi XBB, AY802779 v FJ965342
XBB
AY802779 CYS FJ965342
100
93,3
100
91,1
99,3 100
99,6
99,7 99,6 100
0.4
Hnh 3.7. Hnh cy so snh mc tng ng trnh t nucleotide ca gen DREB1 ging u tng
Hinh 3.8. So snh trinh t amino acid ca CYS, AY802779, FJ965342 v XBB
XBB
AY802779 CYS FJ965342
100
98,3
100
97,0
97,9 100
98,7
99,6 98,3 100
Hnh 3.9. Hnh cy so snh mc tng ng trnh t amino acid ca gen DREB1 ging u tng CYS
- a tch dng gen DREB1 ca hai ging u tng Cuc vng Yn Sn v Vng Ngn Sn
- a xc nh trnh t gen DREB1 ca ging u tng Cuc vng Yn Sn
2. ngh
Cn tip tuc nghin cu v gen DREB1 lm c s thit k vector to cy chuyn gen DREB1 1c kh nng chu hn