2008, 21 (4) : 493 499 Forest Research : 100121498 (2008) 0420493207 7, 3,,, (, 650224) : Giem sa 7, ; 7 :,, : ; ; ; ; : Q996 : A The Chrom osom e Ana lysis of 7 In sect Cell L ines from L ep idoptera ZHANG X in, FENG Ying, D ING W ei2feng, MA Tao, MA Yan (Research Institute of Resource Insects, CAF; The Key Laboratory of Cultivating and U tilization of Resource Insects, State Forestry Adm inistration, Kunm ing 650224, Yunnan, China) Abstract: The chromosomes of 7 insect cell lines from Lep idop tera were analysed. The results showed that the favorable treated concentration and time of colchicines solution and hypotonic KC l solution for each cell line, and the chromosomes of the 7 Lep idop tera cells disp layed the typ ical karyotype characteristics of Lep idop tera cell lines: these cells had a great variation in chromosome num ber, and had deep ly heterop loid, chromosomes w ith diffuse centeromeres were short pole2like, pellet2like and sphere2like. Key words:lep idop tera; cell lines; chromosome; karyotype; heterop loid,, [ 1-2 ] Sm ith [ 3 ],,,, [ 4 ],, [ 5 ] [ 6 ] ;,, DNA, (Lep idop tera) [ 7-9 ] : 2007212212 : 948 ( : 2002252) ( : 4238) : (1981 ),,,. 3.
494 21 [ 10-11 ], 7, Sf9 Sf21,, ; 7 1 1. 1 1. 1. 1 Ae ( A n2 theraea eucalypti Scott ), SES2MaB r22 ( M am estra brassicae L innaeus ), N IAS2MaB r292 ( 5 ), Sf21 ( S podoptera frugiperda Sm ith ), Sf9 ( ), N IAS2Px2 ( Papilio xuthus L innaeus ), FR I2Sp Im 21229 ( Spilosom a im pari2 lis Butler 5 ) Ae SES2MaB r22 N IAS2 MaB r292 N IAS2Px258 FR I2Sp Im 21229 MGM450 + 10% FBS (Hylone) ; Sf21 Sf9 TNM2FH + 10% FBS(Hylone), 25 1. 1. 2 : 0. 005 mol L - 1 ; KCL : 0. 50% 0. 58% 0. 65% ; Carnoy : = 3 1; : 1 /15 mol L - 1 Na 2 HPO 4 1 /15mol L - 1 KH 2 PO 4 4 1 ; Giem sa : 1 Giem sa 10 ; 1. 2 1. 2. 1, 1 /10 0. 005 mol L - 1, 4 h 4. 5 h 5 h 1. 2. 2, 150 g 5 m in, ; 7 ml 0. 50% 0. 58% 0. 65% KCl 15 m in, 150 g 5 m in, 1. 2. 3 1 ml KCl, 1 ml ( ), 6 ml, 15 m in, 150 g 5 m in, 1. 2. 4 7 ml,, 15 m in, 150 g 5 m in, 1. 2. 5 (0. 1 0. 3 ml, ),, 1. 2. 6 10 15 m in,, 1. 2. 7 100, IM50 ; 2 2. 1 Ae Ae, ( 1) : KCl 0. 50%,,, ; KCl 0. 58% 0. 65%,, KCl 0. 50%, 4. 5 h, 5 h, 4 h, Ae 0. 005 mol L - 1 4. 5 h, KCl 0. 50%, 15 m in Ae [ 9, 12 ],,, ;, 2 m ;,, ( 1),, 100, Ae 91, 472, 343, 340 370 ( 1)
4 : 7 495 1 7 0. 005 mol L - 1 /h KCl (15m in) /% Ae SES2MaB r22 N IAS2MaB r292 Sf21 Sf9 N IAS2Px258 FR I2Sp Im21229 4 0. 50 + - + + + + - + 4. 5 0. 50 + + + - + + + + + + + - + + 5 0. 50 + + - + + + + + + - + + + 4 0. 58 - - - + + + - - 4. 5 0. 58 - - - + + + + - - 5 0. 58 - - - + + + + - - 4 0. 65 - - - - + + + + + - 4. 5 0. 65 - + + - - + + + + + - 5 0. 65 - + + + - - + + + - - : + + +, + +, +, - 1 Ae,, ( 2),, 100 134, 243, 104, 110 140 ( 2), 110 150, 2. 2 SES2M abr22 SES2MaB r22, ( 1) : KCl 0. 65%,,, KCl 0. 50% 0. 58%,,, KCl 0. 65%, 5 h ; 4. 5 h ; 4 h,, SES2MaB r22 0. 005 mol L - 1 5 h, 0. 65% KCl 15 m in SES2MaB r22,,, SES2MaB r22, 5 m, 2 SES2MaB r22 2. 3 N IAS2M abr292 N IAS2MaB r292, ( 1) : KCl 0. 50%,,, ; 0. 58% 0. 65% KCl,,,, KCl 0. 50%, 3, 4. 5 h, 5 h, 4 h,
496 21 N IAS2MaB r292 0. 005 mol L - 1 4. 5 h, 0. 50% KCl 15 m in N IAS2MaB r292 ;, ; ; ; 2 m,,, ( 3),, 100, 129, 560, 302, 280 330 ( 3),, Sf21,, ;, 5. 5 m, 2 m,,, ;,, ( 4), 100, 223, 376, 144, 170 200 ( 4) 4 Sf21 2. 4 Sf21 3 N IAS2MaB r292 Sf21, ( 1) : KCl 0. 50% 0. 58%,,, 0. 50% KCL KCl 0. 65%,,,, Sf21 KCl Sf21, KCl 0. 50% 0. 58%, 4 h, Sf21 4. 5 5 h, 0. 50% KCl 15 m in Sf21 2. 5 Sf9 Sf9, ( 1) :, 3, 0. 65% KCl,, 0. 58% 0. 50%,,, KCl 0. 65%, 3, 4 h, Sf9 4. 5 5 h, 0. 65% KCl 15 m in Sf9, Sf21,,,, 2 6 m ;, ;,,
4 : 7 497 ( 5), 100, 186, 402, 142, 160 190 ( 5), Sf9, 6 N IAS2Px258 2. 6 N IAS2Px258 5 Sf9 4 4. 5 5 h KCl 0. 50% 0. 58% 0. 65% 15 m in 9 N IAS2 Px258, ( 1) : KCl 0. 65%,,, KCl 0. 50% 0. 58%, :,, KCl 0. 65%, 4 h ; 4. 5 h ; 5 h,, N IAS2Px258 0. 005 mol L - 1 4 h, 0. 65% KCl 15 m in N IAS2Px258,, 1 2 m ;,,,, ( 6),,, 100 200,, 84, 412, 184 ( 6) 2. 7 FR I2Sp Im 21229 4 4. 5 5h KCl 0. 50% 0. 58% 0. 65% 15 m in 9 FR I2 Sp Im21229, ( 1) : KCl 0. 50%,,, ; KCl 0. 58% 0. 65%, KCl 0. 50%, 5 h, 4. 5 h, 4 h, Ae 5 h, KCl 0. 50%, 15 m in FR I2Sp Im21229,,, ;, ;, 1 5 m,, ( 7) 100 200,, 43, 467, 181 ( 7) 3 :, 7 : ;
498 21 7 FR I2SP IM21229 : 1: Ae, 2: SES2MaB r22, 3: N IAS2MaB r292, 4: Sf21, 5: Sf9, 6: N IAS2Px258, 7: FR I2Sp Im21229,, 7,,,, 9, 7,,,, [ 13 ],,,, 2n = 56 [ 14 ], n = 31 [ 15 ] n = 31 [ 16 ],, 7,, [ 17 ], [ 18 ],, [ 19 ],, [ 20 ] :, ph ; [ 21 ] ; [ 22 ], ; Peter [ 23 ],, :,,,,,
4 : 7 499,,, ;, : [ 1 ],,,. [ J ]., 2004, 17 (1) : 116-124 [ 2 ],,. [ J ]., 2006, 28 (3) : 20-24 [ 3 ] Smith G E. Production of human beta interferon in insect cells infec2 ted with a baculovirus exp ression vector[ J ]. Mol CellB iol, 1983, 13: 183-192 [ 4 ] KarlMaramorosch. : [ J ]., 1992 (4) : 30-32 [ 5 ],,,. BmN [ J ]., 2003, 29 (2) : 136-141 [ 6 ],,,. [ J ]. ( ), 2006, 20 ( 1 ) : 76-77 [ 7 ] Jarman2Sm ith R F, A rm strong S J, A l2rubeaim. Chromosome Insta2 bility in Spodoptera Frugiperda Sf29 CellL ine[ J ]. B iotechnol Prog, 2002, 18: 623-682 [ 8 ],,,. [ J ]., 2002, 45 (2) : 279-282 [ 9 ],. [ J ]., 1999, 36 (4) : 233-238 [ 10 ] Jun M itsushashi. Invertebrate Tissue CultureMethods[M ]. Tokyo: Sp ringer2verlag, 2002: 323 [ 11 ],,,. [ J ]., 2006, 19 (1) : 32-38 [ 12 ] Thom son J A, Grace T D C. Cytological observations on cell strains established in culture from insect Ovarian tissue [ J ]. Aust J B iol Sci, 1963, 16: 869-876 [ 13 ],,,. [ J ]., 2007, 20 (4) : 551-555 [ 14 ],,,. [ J ]., 2006, 28 (9) : 1167-1172 [ 15 ],,,. [ J ]., 2005, 48 (1) : 135-138 [ 16 ],,,. [ J ]., 2003, 46 (6) : 773-782 [ 17 ],,. [ J ]., 2000, 26 (1) : 34-37 [ 18 ],,,. BHS [ J ]., 1993, 12 (4) : 265-270 [ 19 ] W harton J H, Ellender R D, M iddlebrooka B L, et al. Fish cell culture: characteristics of a cell line from the ailver perch, B aird2 iella vhrysura[ J ]. In V itro Cell Dev B iol, 1997, 13 (6) : 389-397 [ 20 ]. [M ]. :, 1986: 30 [ 21 ],,,. HLAMP [M ] / /. :, 1993: 160-167 [ 22 ]. [ J ]., 1994, 16 (1) : 4-14 [ 23 ] Peter Raicu, Francise M ixich. Cytogenetic effects of sodium azide encapsulated in liposomes on Heteroploid cell culture[ J ]. Mutation Research Letter, 1992, 283 (3) : 215-219