Neomycin phosphotransferase II

Maximum residue limit Mass spectrometry Tandem mass spectrometry Material safety data sheet North American Free Trade Act N-Hydroxysuccinimide Nitrogen-phosphorus detection Neomycin phosphotransferase II Optical density Office of Plant Protection and Quarantine... 

Neomycin phosphotransferase II (NPTII) extraction from cotton leaves and cottonseed. The extraction buffer consists of 100 mM Tris, lOmM sodium borate, 5mM magnesium chloride, 0.2% ascorbate and 0.05% Tween 20 at pH 7.8. The frozen leaf sample is homogenized in cold (4 °C) buffer. An aliquot of the homogenate is transferred to a microfuge tube and centrifuged at 12 000 g for 15 min. The supernatant is diluted and assayed directly by ELISA. 

Van Houdt, H. Kovarik, A. Van Montagu, M. Depicker, A. Cross-talk between posttranscriptionally silenced neomycin phosphotransferase II transgenes. FEES Lett, 467, 41-46 (2000)... 

Kromer, W.J. Bailey, J.E. Expression of the membrane protein glycophorin A as a fusion with the antibiotic resistance protein neomycin phosphotransferase II. Biotechnol. Bioeng., 57, 238-244 (1998)... 

Figure I. T-DNA region of pIESIAI andpIJ2B6plasmids used to express human CYPl A1 and CYP2B6 in transgenic rice plants. RB, right border LB, left border NOS, nopaline synthase promoter NT, nopaline synthase terminator NPTII, neomycin phosphotransferase II 35S, cauliflower mosaic virus (CaMV) 35S promoter E7, seven-enhancer region (-290 to -90) from CaMV 35S promoter AMV-5 VTR, alfalfa mosaic virus 5 -untranslated region HPT, hygromycin B phosphotransferase.

The first experiments involving foreign gene expression in higher plants used the coding regions of the bacterial genes for chloramphenicol acetyl transferase (CAT) (47) and neomycin phosphotransferase (NPT-II) (48) fused to the nopallne synthase promoter (49,50). When neomycin phosphotransferase was expressed in tobacco, it was able to detoxify kanamycin, which normally kills plant cells (35,51-53). 

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