23 6 CHINA BIOTECHNOLOGY 2003 6 3 3 3 ( 410128),,,,, [1 ],,,, [1,2 ],,,,, ( kanamycin, Kan) (hygromycin,hyg), (geneticin,g418), 1,, ; :2002212226 :2003203210 3 863 (101201201201) 3 3,:yzlzj @public. cs. hn. gov,, (Kan r ) ( npt2 ), ( aph 2 ), ( E. coli) apha2 (APH(3 ) ) [3] Tn5, : npt2,, ( npt2 ),, 2 2. 1,,,,,
10 23, [3 ],,,,,,,,, 500mgΠL,,, Cho [4 ],, K599, 200mgΠL, [5 ], 75mgΠL, [6 ], 100mgΠL,,, [7 ] ( npt2 ),,,,,,,,, [8,9 ], [10 ],,, 2. 2 2. 2. 1,, [11 ] (T0 ), (T1 ), 3 1,, [12 ] T2, T22T4, T2 F1 F2, 300mgΠL,,, Dubreucq [13 ], T4,,, 2. 2. 2,Lee [14 ],,,, : [15 ],,, [16 ], 300mgΠL,, [17 ] Bt,, 2. 2. 3,, [18 ] 2. 5 %,
6 : 11 [19 ] Bt 750mgΠL,5,,,,, 3, :???? 3. 1 [1 ],,?, 5cm 0113 gπml, 5000 [1 ] :,,,,,, [20 ],,, 3. 2, :,, [21 ], :,Smalla [22 ] npt2, 5000, PCR 3. 3,?,, ;,,,,,, 3. 4,Vries [23 ] npt2 FLAVR2SAVRTM 0. 1 %, APH(3 )2 0. 08 %, 25 74ngΠ (kg d),aph (3 )2,
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6 : 13 The Application of Kanamycin in Transgenic Plants and the Biosafety Assessment of Kan r Gene Wang Zixuan Yi Zili (Hunan Agricultural University Cell Engineering Key Laboratory Changsha 410128) Abstract The functional mechanism and applications of Kanamycin in screening transgenic plants and their progenys were reviewed. Kanamycin could apply to distinguish the transformants from non2transformants during transformation,and to genetic analyses of transformed progenys,or purity assay of seeds,moreover to screen the mature plants of transformed progenys in the field. With the wide application of Kanamycin,its biosafety was more and more emphasized. In this paper,the advances were summarized on the weeding of transgenic plants,horizontal expansion of Kanamycin gene, and its safety for medicine and food. Key words Kanamycin Transgenic plant Biosafety ( 8 ) [ 11 ] Gottesman S, Wickner S, Maurizi MR. Protein quality control : triage by chaperones and proteases. Genes Dev,1997,11 :815 823 [ 12 ] Park J, Ryu J, Kim KA, et al. Mutational analysis of a human immunodeficiency virus type 1 Tat protein transduction domain which is required for delivery of an exogenous protein into mammalian cells. J Gen Virol, 2002,83 :1173 1181 [ 13 ] Ho A, Schwarze SR, Mermelstein SJ, et al. Synthetic protein transduction domains : enhanced transduction potential in vitro and in vivo. : Cancer Res, 2001,61 : 474 477 [ 14 ] Jin LH, Bahn JH, Eum WS,et al. Transduction of human catalase mediated by an HIV21 TAT protein basic domain and arginine2rich peptides into mammalian cells. Free Radic Biol Med 2001,31 :1509 1519 [ 15 ] Kim DT, Mitchell DJ, Brockstedt DG,et al. Introduction of soluble proteins into the MHC class I pathway by conjugation to an HIV tat peptide. J Immunol, 1997,159 :1666 1668 [ 16 ] Vocero2Akbani A, Chellaiah MA, Hruska KA,et al. Killing HIV2 effected cells by transduction with an caspase23 protein. Nat Med, 1999,5, 29 33 HIV protease2activated [ 17 ] Cao G, Pei W, Ge H,et al. In Vivo Delivery of a Bcl2xL fusion protein containing the TAT protein transduction domain protects against ischemic brain injury and neuronal apoptosis. J Neurosci, 2002,22 :5423 5431 Progress of HIV2TatPTD in Medicine Ding Jin Ma Bin Liu Jun Xue Caifang (Department of Etiology The Fourth Military Medical University Xi an 710032) Abstract Compounds, peptides or DNA linked covalently with HIV2TatPTD could enter cells in a receptor and transporter2independent fashion. Some of them can even cross the blood2brain barrier. Up to date, the exact mechanism in which the TatPTD is able to target and to transverse lipid membranes remains unknown. Characterized with prominent transduction activity, TatPTD has been widely applied in medical research, and would be a potent tool for the treatment of human disease. Key words Protein transduction domain (PTD) HIV2Tat