Tách dòng cDNA và thiết kế vector biểu hiện thụ thể neurokinin - 1

Tap chi Cong nghe Sinh hpc 11(4): 653-658, 2013

TACH DONG cDNA VA THIET KE VECTOR BIEU HIEN THU THE NEUROKININ-1

V5 Thj T hinm g L an1, Le H ong T hu1, Dinh Do&n L ong1,2

'Truang Dai hpc Khoa hpc tu nhien2, Dai hpc Quoc Gia Ha Noi

2Khoa Y-Duprc, Dai hpc Quoc Gia Ha Noi

TOM TAT

Thu the neurokinin-1 (NK1-R) thupc nhom cac thu the xuyen mang lien ket vai G protein co vai tro quyet

dinh hoat dong cua cac te bao than kinh, tham gia vao viec dieu hoa cac phan xa non, ho hap va cac hanh vi

phan xa. Thu the NK1-R duoc phien ma tir gen dan ban nhung gom hai dang sai khac nhau a dau carboxyl

hinh thanh do cat noi luan phien mRNA. Hai dang NK1 -R co ai lire lien ket khac nhau vcri cac chat dong van

hoac cac chat doi van. Bieu hien NK1-R tai to hop trong he thong te bao dong vat la phucmg phap toi uu

hien nay de sang loc cac chat doi van/dong van canh tranh voi phoi tu trong san xuat cac biet duoc dieu tri cac

benh than kinh, roi loan dap ung mien djch. Trong bai bao nay, chung toi trinh bay ket qua phan lap cDNA

hoan chinh ma cho NK1-R tu mo phoi cua nguai Viet Nam. Trinh tu nucleotide va trinh tu amino acid suy

dien cua NK1-R phan lap tir mo phoi giong vai NK1-R phan lap tir mo nao ciia ngudi Viet va co 4 amino acid

sai khac so vai trinh tu goc trong ngan hang du lieu (Databank). Tuy nhien, phan tich in silico cho thay 4 sai

khac nay khong anh hudng den cau true xuyen mang ciia thu the. cDNA hoan chinh duac dua vao vector bieu

hien pEGFP-Nl nham muc dich xay dung he thong te bao bieu hien NK1-R tai to hop de sang loc cac chat

dong van va doi van thu the tir nguon duac lieu Viet Nam.

Tir khoa: Chat doi van thu the, chat dong van thu the, phoi tic P, thu the neurokinin-1 (NK1-R), thu the xuyen

mang lien ket G protein.

MO DAU

Thu the neurokinin-1 (N K 1-R) la thanh vien

trong ho thu the chira 7 viing ky nude xuyen mang.

Dau carboxyl cua N K 1-R lien ket vdi G protein ben

trong te bao chat; dau am ino lien ket chu yeu vdi

chat P (phoi tir) d ben ngoai mang te bao (Severini et

al., 2000). N K1-R bat dau con dudng truyen tai cac

tin hieu gay dau ddn va dap irng sinh li (ho hap, dieu

hoa, cac hanh vi phan xa) cho te bao (InHae, Tae,

1995). C) ngudi, gen ma hoa N K 1-R la gen don ban

co 5 exon, nam tren nhiem sac the so 2. Sau phien

ma, co 2 dang m RN A (khac nhau d viing 3 ’) duoc

djch ma tong hop hai dang N K 1-R co ai lire khac

nhau vdi cac chat dong van hoac cac chat doi van thu

the (Jian et al., 2008). Trong khi cac chat dong van

lien ket vdi thu the va co tac dung tuong tir nhu cac

phoi tu (ligands) thi cac chat doi van lien ket vdi thu

the nhung khong gay ra cac phan irng sinh hoc hoac

lam giam hay khoa cac phan ung trung gian cua chat

dong van (Rosso, M u'noz, 2006). Phat trien cac

thuoc dong van hay doi van N K 1-R dac biet co y

nghTa doi vdi dieu tri nhieu benh, vi du nhu thudc

giam dau, chong lo au. tram cam, chua cac benh

dudng ho hap, dieu trj chim g buon non va sir non

m ua do tac dung cua hoa tri lieu ung thu (Cham indi

et al., 2009; Rogers, Blackburn, 2010). M ot img

dung rong rai cua thuoc doi van N K 1-R la tham gia

vao viec dieu trj chim g tam than phan liet, tram cam

co lien quan den viec tang lugng dopam ine trong nao

bo (Rupnipak, Kram er, 2002). Khi phoi tir P lien ket

vdi N K 1-R thi lugng dopam ine tang. Co the dieu

chinh lugng dopam ine bang cach sir dung cac thuoc

doi van lien ket vdi N K 1-R nham ngan chan sir

tuong tac ciia chat P (Rupnipak, Kram er, 2002).

N am 2003, lan dau tien chat doi van N K 1-R la

aprepitant (Em end) dugc hiep hoi quan ly thuoc va

thuc pham (FDA) Hoa Ky phe chuan cho phep san

xuat va luu thong tren thj trudng (Rogers,

Blackburn, 2010).

Viec tim kiem cac chat doi van hoac dong van

N K 1-R hau het dua vao he thong te bao bieu hien

N K 1-R noi sinh hoac N K 1-R tai to hgp (Q uartara,

Altam ura, 2006). Vdi uu diem noi bat ve so lugng va

phuong thirc bieu hien nen N K 1-R tai to hgp dugc

sir dung trong cac nghien ciru cau true, dong hoc

tuong tac ciia N K 1-R vdi chat dong van hoac chu

van (tang kha nang phat hien tucmg tac, chu dong

gay dot bien dinh hudng lam thay doi cau true NK1 -

R ,...) (Q uartara, Altam ura, 2006). Vi vay, phan lap

cac cD N A hoan chinh, bieu hien N K 1-R tai to hgp;

tir do thiet lap cac he thong te bao sang loc thudc tir

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