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从小麦品种“Ｂｏｄａｌｉｎ”胚性悬浮细胞分离出原生质体，通过电激将质粒ＰＢＣ１ＤＮＡ（携带β－葡萄糖苷酸酶（ＧＵＳ）标记基因和潮霉素抗性基因ｈｐｈ）导入原生质体。采用ＢＴＸ电激系统和ＡＳＰ电激缓冲液，最佳电激条件为３００Ｖ（７５０Ｖ／ｃｍ）和５０ｍｓ（约１０００μＦ），转化的原生质体内ＧＵＳ的活性最高；质粒ＤＮＡ的有效使用浓度为２５μｇ／ｍｌ。电激处理后，原生质体培养２～３天，ＧＵＳ基因表达最强，宜于检测其瞬时表达；牛胸腺ＤＮＡ可协助提高ＧＵＳ基因的导入效果。质粒ＰＢＣ１ＤＮＡ处理的原生质体培养于添加潮霉素的ＫＭＰ培养基。经４个月抗性筛选，选择获得１５个潮霉素抗性克隆
Protoplasts were isolated from embryogenic suspension cells of the wheat variety “Bodalin” and the plasmid PBC1 DNA (carrying the beta-glucuronidase (GUS) marker gene and the hygromycin resistance gene hph) was introduced into the protoplasts by electroporation. The best electroporation conditions were 300V (750V / cm) and 50ms (about 1000μF) using BTX system and ASP buffer. The highest activity of GUS in transformed protoplasts was obtained. The effective concentration of plasmid DNA was 25μg / ml . After electroporation, the protoplasts cultured for 2 to 3 days, GUS gene expression is strongest, suitable for detecting transient expression; bovine thymus DNA can help improve the GUS gene introduction effect. Plasmid PBC1 DNA-treated protoplasts were cultured in hygromycin-supplemented KMP medium. After 4 months resistance screening, 15 hygromycin resistant clones were selected