(classical sw ine fever, CSF) (CSFV ) (O IE) A 15, ,, C2,, , 1. 1 SXZZ), 1. 2

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1 31 5 () V o l. 31 N o Jour. of N o rthw est Sci2T ech U niv. of A gri. and Fo r. (N aṫ Sci. Ed. ) O cṫ 2003 E2 gp55 Ξ 1, 1, 1, 1, 1, 2, 2 (1, ; 2, ) [ ] R T 2PCR PCR E 2,, A lfo rt B rescia C2, ; DNA star E 2 gp 55, (C2),, gp 55, C [ ] ; E 2 ; gp 55 ; ; ; []S []A [] (2003) (classical sw ine fever, CSF) (CSFV ) (O IE) A 15, ,, C2,, , [1 ] (F laviviridae) (P estiv irus), RNA, kb, (OR F ), 11 [2, 3 ], E 2 gp 55,, gp 55 gp 55, E 2 [4, 5 ] gp 55 gp 55,, (C2) gp 55,, ( SXZZ), 1. 2 (AM V ) T aqdna PCR RNA PM D 218T () ; DNA BO EHR IN GER M ANN H E IM ; DNA star () 1. 3 CSFV A lfo rt [2 ] B rescia [3 ] C2 [6 ], 4, (), (C2) E RNA R T 2PCR npcr R T 2PCR 20 ΛL, 42 1 h; PCR 50 Ξ [] [](2001K2022G922) ; ( ) [](1976- ),,,, [](1956- ),,,,,,

2 5 : E 2 gp ΛL, 98 8 m in, s, 50 s, s, s, 35, s, s, 35, m in, PCR m in; npcr : 98 8 m in, 98 1 T able 1T he code, sequence, location and length of the p rim ers P rim er E 1 E1 E 2 E2 Sequence L ocation gnt L ength 5 T GTA T TA GA CCA GA CT GG CT GCCAA CCGCCGTCTA TCT T CGGCTA GCCT GTAA GGAA GA T GGTCGCCGCTCAA CAA GA CA E 2 E 2 PM D 218T,, PCR, () 1. 6E 2 gp 55 DNA star Jam eson2w o lf gp E 2 gp 55 DNA star Kyte2Doo little gp E 2 gp 55 DNA star Gam ier2rob son gp E 2 E 2E 2 npcr, npcr PGEM 2T easy PCR 1 1, npcr (1 119 bp ), PCR (1 119 bp ) Strain 1nPCR PCR M. DL 2000; 1. E 2, E 2 npcr ; 2. ; 3. PCR F ig. 1nPCR p roduct, PCR and restriction analysis of recom binant p lasm id M. DNA m arker DL 2000; 1. PCR p roduct amp lified w ith p rim ers E 2 and E2 ; 2. Restriction analysis of recom binant p lasm id; 3. PCR identification of recom binant p lasm id 2. 2gp 55 E 2 gp 55, E 2 gp 55 gp gp 55 T able 2Comparison of the am ino2acid sequence gp 55 p ro tein of CSFV Homo logy of the am ino2acid A lfo rt B rescia C SXZZ GXYR HN KF Shim en A lfo rt B rescia C SXZZ GXYR HN KF 86. 3

3 130 () 31 (SXZZ ) gp % (GXYR ) [7 ] 97. 3% (HN KF) [8 ], (C2) 86. 9%, 88. 7% ( Sh im en ) [9 ], 89. 5% (A lfo rt ) [2 ] 90. 3% (B rescia) [3 ] gp 55,, 2. 3 E 2 gp 55 gp 55,,, gp 55, gp 55 A B C D 4, 4 gp 55 N , C ,, gp 55 A , A 3, A 1 A 2 A 3; B ; C ; D ,A 1 B C,,A 2 A 3 D, [4, 5 ] DNA star Jam eson2w o lf gp 55, E 2 gp 55 (2, 1 gp 55 N 690 ) 2, (C2) , , , , , , (SXZZ ) , , , , , , , , C2 gp ,, 2E 2 gp 55 F ig. 2A ntigenicity of E 2 gene code gp 55 p ro tein of CSFV 2. 4E 2 gp 55 gp 55 [10, 11 ] DNA star Kyte2Doo little, gp 55 gp 55 gp 33, E 2 gp 55 3 (1 gp 55 C 40 gp 55 N 690 ) 3, ( tran sm em b rane region, (C2) (SXZZ )C TM R ), 18, 3E 2 gp 55 F ig. 3H ydrophobicity of E 2 gene code gp 55 p ro tein of CSFV

4 5 : E 2 gp E 2 gp 55 W en svoo rt G [12 ] 1989 gp 55,, gp 55 2, B C, A D 2, DNA star Gam ier2rob son gp 55, E 2 gp 55 4 ( 1 gp 55 N 690 ) 4, gp 55 ΑΒ, (C2) (SXZZ ), 3 4E 2 gp 55 F ig. 4Secondary structure of E 2 gene code gp 55 p ro tein of CSFV,,,,,, Jones [13 ] Jam eson2w o lf M SA 2 (51 ku), D eleage [14 ] 59, 72%,, 65%,, F ro shauer [15 ] Kyte2Doo little m al F, 23, 2,,, gp 55, gp 55, gp 55, gp 55, gp 55, C [] [ 1 ],,,. [A ].,. [C ]. :, [ 2 ]M eyers G, Rum enapf T, T h ie H. M o lecular cloning and nucleo tide sequence of hog cho lera virus[j ]. V iro logy, 1989, 171: [ 3 ]M oo rm ann R J M,W arm erdam P M, H ulst M M, et al. M o lecular cloning and nucleo tide sequence of hog cho lera virus strain B rescia and m app ing of the genom e region encoding envelope p ro tein E1[J ]. V iro logy, 1990, 177: [ 4 ]V an R ijin P A. A p relim inary m ap of ep itopes on envelope glycop ro tein E1 of HCV strain B rescia [J ]. V eterinary M icrobio logy, 1992, 33: [ 5 ]V an R ijin P A. A ntigen structure of envelope glycop ro tein E1 of hog cho lera virus[j ]. Journal of General V iro logy, 1994, 68: [ 6 ],,,. (HCLV ) gp55 p54 p80 p54

5 132 () 31 [J ]., 1998, 28 (6) : [ 7 ],,,. C2gp55 (E2) DNA [J ]. ( ), 2002, 30 (1) : [8 ],,,. E2 [J ]. (), 2002, 30 (2) : [ 9 ]. [D ]. :, [ 10 ]H ulst M M, Pano to F E, Hoekm an A, et al. Inactivation of the RN ase activity of glycop ro tein E rns of classical sw ine fever virus results in a cytopathogenic virus[j ]. J V iro l, 1998, 72 (1) : [ 11 ]W ang C, Sarnow P, SiddiguiA. T ranslation of hum an hepatitis C virus RNA in cultured cells is m ediated by a internal ribsom e binding m echanism [J ]. J V iro l, 1993, 67: [ 12 ]W ensvoo rt G. Topograph ical and functional m app ing of ep itopes on hog cho lera virus w ith monoclonal antibodies[j ]. J Gen V iro l, 1989, 70: [ 13 ]Jones G L, Edm undson H M, L o rd R, et al. Imm uno logical fine structure of the variable and constant regions of a po lymo rph ic m alarial surface antigen from P lasm od ium f a lcip a rum [J ]. M o l B iochem Parasito l, 1991, 48 (1) : 1-9. [ 14 ]D eleage G, Roux B. A n algo rithm fo r p ro tein secondary structure p rediction based on class p rediction[j ]. P ro tein Eng, 1987, 1 (4) : [ 15 ]F ro shauer S, Beckw ith J. T he nucleo tide sequence of the gene fo r m alf p ro tein, an inner m em brane component of the m alto se transpo rt system of E scherich ia coli[j ]. J B io l Chem, 1984, 259 (17) : Sim u lation analysis of E 2 gene code gp 55 p ro tein of classical sw ine fever viru s ZHANG Y ong-guo 1, ZHANG Yan-m ing 1, X ING Fu- shan 1, XU X in-gang 1, HU J ian-he 1,L IU X iang- tao 2, HAN Xue-q ing 2 (1 College of A nim al S cience and T echnology,n orthw est S ci2t ech U niversity of A g riculture and Forestry, Yang ling, S haanx i , China; 2 L anzhou V eterinary R esearch Institute, Chinese A cademy of S ciences,l anzhou, Gansu , China) Abstract: T he E 2 gene of p revalen t viru len t strain of classical sw ine fever viru s w as amp lified by the R T 2PCR and PCR. T he amp lified fragm en ts w as cloned in to vecto r and then sequenced, based on the am ino - acid sequences of A lfo rt and B rescia strain s, am ino2acid sequence of gp 55 p ro tein w ere deduced. T he resu lts show ed that there w as som e differences among recen tly p revalen t viru len t strain s, C2strain and viru len t strain ṡ A n tigen icity, hydrophob icity and secondary structu re of E 2 gene code gp 55 p ro tein of classical sw ine fever viru s w ere analysed u sing DNA star soft w are. T he p revalen t viru len t strain and C2 strain an tigen icity, hydrophob icity and secondary structu re w ere compared. T he resu lts show ed that an tigen icity and secondary structu re w ere differen t betw een the p revalen t viru len t strain and C2strain, the C2term inal hydrophob icity has little difference betw een the p revalen t viru len t strain and C2strain. Key words: classical sw ine fever viru s; E 2 gene; gp 55 p ro tein; an tigen icity; hydrophob icity; secondary structu re