Biểu hiện gen HIV-1-P24 trong cây cà chua (Lycopersicon esculentum Mill.) chuyển gen vào lục lạp

Tgp chi Cong nghe Sinh hgc 12(2): 281-288, 2014

BIEU HIEN GEN HIV'1-P24 TRONG CA Y C A CHU A {LYCOPERSICON ESCULENTVM

Mill.) CHUYE N GEN VAO Ly C LAP

Pham Dire Tri, Nguyen Thi Thanh, Nguyen HChi Ho

Vien Sinh hgc nhi^t d&i, Vien Hdn ldm Khoa hgc vd Cong nghi Viet Nam

Ngiy nhin bai'09.5.2014

Ngiy nhin ding:19.6.20I4

TOM TA T

Ciing vdi cdng nghg chuyen gen vao bO gen nhin, cdng nghe chuyen gen vao luc lap dang duoc iing dung

d6 tgo nhOng th\rc vat cd kha nang san xuat protein tai to hgp ung dung trong y duac. Diing phuong phap ban

gen vio li that va la mam, de chuyen gen HiV-i-p24 vao luc lap ca chua giong Trang Ndng 386 Mau sau hSn

gen dirge d$t trgn mfii tmdng tgo md seo co klioang MS vitamin B5 lmg/1 BA, 0,lmg/l NAA co spetinomycin.

T3ng din ap Igc chgn lgc bang khang sinh spectinomycin tir 50mg/I len 300mg/l de cd the nhan dugc mo s?o

chuyen gen dong lap the. Tai sinh cay ca chua tir mo seo chuyen gen bang moi trudng MS vitamin B5 cd bo

sung 2,0mg/l zeatin, 0,2mg/l lAA, 300mg/l spectinomycin. Nh^n dugc b6n ddng cay c^ chua chuyen gen ddng

lap the dugc mmh chiing bSng ky thuat PCR va Southem blot. Trong bon dong nay, ba dong tir mau la that va

mdt ddng tii mau la mam. Protein tai to hgp HIV-l-p24 trong cay ca chua chuyen gen dugc dinh tinh bang

Western blot va dinh lugng bang phuong phap ELISA, Ket qua thu diroc cay ca chua chuyen gen cd h^m

lugng protein HlV-1 -p24 tren protein tong a la dat tmng binh 2,35%. Nhu vSy, da hoan chinh quy trinh chuyen

gen vio luc lgp ci chua bing phuong phap ban gen va cac dong cay ca chua chuyen gen dong lap the deu bieu

h\^agmHIV-l-p24.

Tir khoa: Chuyen gien vdo lye lgp, Gen HIV'I-p24, Khdng nguyen HIV-I-p24. Lycopersicon esculentum Mill,

Spectinomycin

M6DAU

Tir nhiing nam cudi the ky 20, hudng chuyen

gen vao lye lap ciy trdng d i dugc cac nhi khoa hgc

quan tim vi nhirng thuan lgi ciia nd. (Obembe et al,

2010; Wang el al, 2009) di tdng hgp: gen luc lap di

truyen theo me sg h^n che su phit tin gen; sd lugng

bg gen trong luc lap khoing 10.000, se giiip cho gen

muc tieu bieu hien d miic cao hon khi chuyen gen

vao nhin; chuyen gen vao luc lap dugc thuc hien

theo nguyen tic tai td hgp tuang ddng giifa DNA

vector vi DNA luc lap, dieu nay cho phep biet dugc

chinh xie vi tri gen dugc chuyen vio vi khdng con

lo ling higu qua bieu hien, Ndi each khac, chuyen

gen luc l^p khdng cd hien tugng gen im ling nhu

trong chuyen gen nhin.

Cong ngh$ chuyen gen luc lap cdn dugc iing

dyng de tgo cac protein dugc lieu tai to hgp trong thuc

vit. Ben cgnh dd, cie nhi khoa hgc dang dinh hudng

nghien ciiu tao vaccine in dugc tir thyc vit vi nhflng

lgi the tihu: vaccine tir thgc vit cd lgi the li bd qua

cdng dogn len men, tinh sgch, trii lanh, tiem vo trung

nhu vaccine thdng thudng; vaccine khang nguyen

biSu hign trong lgc lgp dugc bio ve khdi sy phan huy

ciia acid v i enzyme trong bao tir, va dugc phdng thich

trong mgt khi vach te bao luc lap bi phin huy bdi

nhung VI khuin trong rudt (Obembe et al., 2010).

Vdi viec tiep can ky thugt hien dgi trong nghien

ciiu chuyen gen luc lap, nghien cim chuyen gen tao

protein khing nguyen HIV-l-p24 vio luc lap cay ca

chua dugc thuc hien. Trong nghien cuu nay, gen

HIV-l-p24 dugc chuyen vio luc lap ciy ci chua

bang ky thuit ban gen dua tren nhung cdng trinh cua

(Apel Wiebke, Bock Ralph, 2009; Ruf el al, 2001;

Wurbs et al, 2007; Zhou et al., 2008).

PHU'ONG PHAP NGHIEN CUV

Gidng c i chua dtrgc dung trong nghien cim la

gidng thuang mai cua cong ty gidng ciy trdng Trang

Ndng: 386. Sii dung la that vi Ii mim cay ca chua

lam vat lieu cho cie nghiefi cim chuyen gen.

Vector chuyen gen luc lap pITB-HIV-l-p24 do

TS Nguyen Thi Thanh thigt kg (Nguydn Thi Thanh

etal, 2010).

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