Guerineau F, Brooks L, Meadows J, Lucy A, Robinson C, Mullineaux P
John Innes Institute, Institute of Plant Science Research, Norwich, UK.
Plant Mol Biol. 1990 Jul;15(1):127-36. doi: 10.1007/BF00017730.
The sulfonamide resistance gene from plasmid R46 encodes for a mutated dihydropteroate synthase insensitive to inhibition by sulfonamides. Its coding sequence was fused to the pea ribulose bisphosphate carboxylase/oxygenase transit peptide sequence. Incubation of isolated chloroplasts with the fusion protein synthesised in vitro, showed that the bacterial enzyme was transported to the chloroplast stroma and processed into a mature form. Expression of the gene fusion in transgenic plants resulted in a high level of resistance to sulfonamides. Direct selection of transformed shoots on leaf explants was efficient using sulfonamides as sole selective agents. Transformed shoots rooted normally on sulfonamides at concentrations toxic for untransformed ones. Sulfonamide resistance was transmitted to the progeny of transformed plants as a single Mendelian dominant character. These results demonstrate that this chimeric gene can be used as an efficient and versatile selectable marker for plant transformation.
来自质粒R46的磺胺抗性基因编码一种对磺胺抑制不敏感的突变二氢蝶酸合酶。其编码序列与豌豆核酮糖二磷酸羧化酶/加氧酶转运肽序列融合。用体外合成的融合蛋白孵育分离的叶绿体,结果表明该细菌酶被转运到叶绿体基质并加工成成熟形式。该基因融合体在转基因植物中的表达导致对磺胺的高度抗性。以磺胺作为唯一选择剂,在叶片外植体上直接筛选转化芽是有效的。转化芽在对未转化芽有毒的浓度的磺胺上能正常生根。磺胺抗性作为单一孟德尔显性性状传递给转化植物的后代。这些结果表明,这种嵌合基因可作为植物转化的一种高效且通用的选择标记。