60 s 72, 60 s 94, 30 s; 5

HTML
DOWNLOAD

Μέγεθος: px

Εμφάνιση ξεκινά από τη σελίδα:

Download "60 s 72, 60 s 94, 30 s; 5"

Τρύφαινα Ζάππας
6 χρόνια πριν
Προβολές:

1 342 Ch inese Journal of Parasito logy and Parasitic D iseases :, CS : PCR CS, M 13, 3 M 132CSP DNA, PCGEN E CS : (PFD - 3gYN ) T 4 W ellcom e N F54 3D 7 7G8 CS, P. f T hgt c : CS (circum spo rozo ite p ro2 tein, CSP), T B, [1-3 ], CS, PCR (PFD - 3gYN )CS, T aq dn T P Sigm a, D YEPR IM ER AB I, T 4DNA Boeh ringer M annheim, X2gal IPT GK Sigm a M 13 R FDNA, T G1 3 : (PFD - 3gYN ), T rager2jen sen [4 ], 70%, 0115%,, CS CS, ( 1), 5 EcoR g, 3 Salg : CSP- P1 5 2GGGAA T TCAA TA GTA G A TCA CT T 2GGA GAA 23, CSP- P2 5 2GG GTCGA CA GT TA CA CTA CA T GGGGA CC A 23 DNA Sethabu tr [5 ] CS, 100 Λl PCR, 25 pmo lg100 Λl, DNA 200 ng, dn T P 200 Λmo lgl 95 5 m in, T aq 2 u PCR : 50, 60 s 72, 60 s 94, 30 s; 5

343 60, 60 s 72, 60 s 94, 30 s, 25 017 % ) CS 1 CS CS F ig11 The CS prote in structure and amplif ied reg ion of the CS prote in partial gene fragmen ṫ CS CS -,, 017%, CS EcoR g + Salg, M 13 mp 18,

2 343 60, 60 s 72, 60 s 94, 30 s, % ) CS 1 CS CS F ig11 The CS prote in structure and amplif ied reg ion of the CS prote in partial gene fragmen ṫ CS CS -,, 017%, CS EcoR g + Salg, M 13 mp 18, 10 Λl, 3 u T 4DNA, 5: 1, ng T G1, X - galg IPT G, PCR CS [6 ] 3 DNA, M 13- CSP DNA D yep rim er PCR PCR, 16%, AB I 373A DNA ( ) PCGEN E CS, CS T 4 ( ) W ellcom e ( ) 3D 7 ( ) N F54 ( ) 7G8 ( CS (PFD - 3gYN ) CS, 830 bp CS R FDNA 811 kb, PCR 830 bp, EcoR g Salg 830 bp ( 2) 2 PCR M 13- CSP, 017% 1 2: M 13- CSP (EcoRg Salg ) ; 3: ΚD NAgEcoR g + H indg markers; 4: PCR markers; 5: CS ; 6 7: M 13- CSP CS. F ig12 The recombinan t bacter iophagesm 13-CSP were identif ied by PCR and restr iction endonucleases and electrophoretic analysis on 017% agarose gel. Lanes 1 2: Two M 13-CSP were digested with EcoRg and Salg respectively; Lane 3: ΚD NAgEcoRg + H indg markers; Lane 4: PCR markers; Lane 5: CS prote in gene fragmen t; Lanes 6 7: PCR-amplif ied products of M 13- CSP cloneṡ D NA 1 CS C (PFD - 3gYN )CS bp, 167, 4017%,

3 % 1716%, : A sp 2A la2a sp 2P ro 2 CS C,, Β CS, T 4 ( ),, 152 ( CS 330 ) 10 ; W ellcom e ( ), T 4,, 30, T 4 W ellcom e,,, 3 F ig13 The homology analysis of the partial am ino ac id sequences of the CS prote in in six P lasm odium f a lcipa rum isolateṡ P. f a lcipa rum ThgTc epitope is underlined. W ellcom e ; 7G8 ( ) 7 17 ; 3D 7 ( ) 8 24 ; N F54 ( ) 6 22 ( 3) 3 PfT hgt c T PFD - 3gYN CS PfT hgt c T Q YL KE IQN SL ST EW SPCSV T,, PFD 23gYN T,, 152 ( CS 330 ), PfT hgt c T CS,, : ; ; 5 ; ; [7-9 ] PFD - 3gYN ( ) CS, 3D 7 W ellcom e N F54 T 4 7G8, PFD - 3gYN CS :, (A sn), (A la) (P ro),, PFD - 3gYN CS A sn A T T A la GCA P ro CCC, CCC PCGEN E, CS

4 345, PFD - 3gYN T 4 ( ),, W ellcom e ( ) 30,,, CS PFD - 3gYN T 4 W ellcom e,, W ellcom e CS, CS W ellcom e CS T B,B, T CS N C CS T, P. f T hgt c T T T, CS T, CS , M o reno (1993) [10 ] CS P. f T hgt c, 5, M HC T, 332 (Q ) (K) (R ), T, (N K) (KT ), T CS, Q YL KE IQN SL ST EW SPCSV T, 332 Q,, T, (KE),, N K KT T, N K KT KE T, 1 N ussenzw eig V and N ussenzw eig RS. C ircum spo ro2 zo ite p ro teins of m alaria parasites. Cell 1985; D el Giudice G, Tougne C, L ouis JF, et al. A m ultip le antigen pep tide from the repetitive sequence of the P lasm od ium m alariae circum spo rozo ite p ro tein in2 duces a specific antibody response in m ice of various H - 2 hap lo types. Eur J Imm uno l 1990; M alik A, Egan JE, Hough ten RA, et al. H um an cyto2 toxic T lymphocytes against the P lasm od ium f alci2 p arum circum spo rozo ite p ro tein. P roc N atl A cad Sci U SA 1991; T rager W and Jensen JB. H um an m alaria parasites in continuous culture. Science 1976; Sethabutr O, B row n A E, Panyim S, et al. D etection of P lasm od ium f alcip arum by po lym erase chain reaction in a field study. J Inf D is 1992; J., E. T., T..... :, 1992: L ockyer M J, Schw arz RT. Strain variation in the cir2 cum spo rozo ite p ro tein gene of P lasm od ium f alci2 p arum. M o l B iochem Parasito l 1987; Caspers P, Gentz R,M atile H, et al. T he circum spo ro2 zo ite p ro tein gene from N F54, a P lasm od ium f alci2 p arum iso late used in m alaria vaccine trials. M o l B iochem Parasito l 1989; C ruz V F, L al A F and M ccutchan T F. Sequence varia2 tion in putative functional dom ains of the circum spo ro2 zo ite p ro tein of P lasm od ium f alcip arum. J B io l Chem 1987; M o reno A, C lavijo P, Edelm an R, et al. CD + 4 T cell clones obtained from P lasm od ium f alcip arum spo ro2 zo ite - imm unized vo lunteers recognize po lymo rphic sequences of the circum spo rozo ite p ro tein. J Imm uno l 1993; ( : )

5 346 Ch inese Journal of Parasito logy and Parasitic D iseases CLON ING AND SEQUENCE ANALY SIS OF THE GENE ENCOD ING THE PART IAL REGION CS PROTE IN OF A PLASM OD IUM FAL C IPARUM ISOLATE FROM Y UNNAN X iao J ianhua 3,L im ing, Chu i Dong,B i H u ix iang,w ang P ing,l i Y ingjie Institu te of T rop ica l M ed icine, T he F irst M ilita ry M ed ical U niversity, Guang z hou ABSTRACT A IM : D eterm in ing nucleo tide sequence of the circum spo rozo ite p ro tein partial gene of the P lasm od ium f a lcip a rum PFD - 3gYN (Yunnan of Ch ina) and finding ou t the differences of the CS gene sequence betw een Ch inese P lasm od ium f a lcip a rum iso late and o ther iso lates. M ETHOD S: T he circum spo rozo ite p ro tein gene fragm en t w as amp lified by po lym erase chain reaction and cloned in to M 13 bacteriophage. M 132CSP single strand DNA s of the th ree po si2 tive clones w ere ex tracted respectively. T hen, the nucleo tide sequence of the CS gene ragm en t w as determ ined by the dideoxy chain term ination m ethod. PCGEN E softw are w as u sed to compare and analyze the CS gene sequence of the six iso lates. RESUL TS: D ifferen t degrees of diversity of the CS gene sequences w ere found among P. f a lcip a rum PFD - 3gYN and o ther iso lates (T 4,W ellcom e,n F54, 3D 7 and 7G8). A non2silen t sub stitu tion at the nucleo tide level being found in the P. f T hgt c an tigen ic ep itope region. CONCL USION: T here w ere differ2 ences in the CS gene sequence among P. f a lcip a rum PFD - 3gYN and tho se of o ther iso lates. Key words: P lasm od ium f a lcip a rum, circum spo rozo ite p ro tein gene, gene sequencing 3 P resent address: D epartm ent of M icrobio logy, H engyang M edical Co llege, H engyang, H unan , 10, 115 kg- 3 kg,, 30 m in- 50 m in 10 d,,,,,, ,,, S arcop tes scabiei [1 ] 015% (50% ),, 5 d 1, 1 7 d 1, 2, :, ( : )

Σχετικά έγγραφα

(HR P ) CD 4 + ) L yt2 ( CD 8 + ) F ITC

(HR P ) CD 4 + ) L yt2 ( CD 8 + ) F ITC 1999; 17 (6) 325329 Ch inese Journal of Parasito logy and Parasitic D iseases 325 FCC1gHN CSP 3 33 510089 : pcdna 3, (H el a ) (CSP), BALBgc : PfCSP H el a,, EL ISAW estern blotting T N K T, BALBgc : EL