水稻寡肽转运基因OsPTR1～10酵母异源表达载体的构建及转化

Construction and Transformation of Heterogeneous Expression Vector in Yeast of Oligopeptide Transport Genes OsPTR1-10 from Rice

[目的]研究水稻中的10个寡肽转运同源基因的功能。[方法]将10个OsPTR基因的开放读码框插入到酵母异源表达载体pDR196中,并将它们转入寡肽转运功能缺失的酵母突变体中,进而测试这些基因的寡肽转运功能。[结果]实际酶切结果与预期酶切结果完全一致,说明目的基因已经成功克隆到酵母穿梭表达载体上。在酵母菌株的营养缺陷型测试中,完全培养基中的所有菌株生长良好;菌株ABC154和ABC738在缺乏Ura、Leu、Lys、His、Trp或Ade的培养基中不能生长,说明菌株ABC738和ABC154是Ura、Leu、Lys、His、Trp和Ade的营养缺陷型菌株,可以用于测试目的基因的寡肽转运功能。菌株pDR196/OsPTR1～10/ABC738和pDR196/ABC738在缺乏Ura的培养基中生长良好,说明质粒pDR196上有Ura基因。[结论]该试验为寡肽转运基因的研究提供了方法支持。

[ Objective ] The purpose was to study the function of 10 homologous oligopeptide transport genes in rice. [ Method ] The open reading frames of the 10 OsPTR genes were inserted into the heterogeneous expression vector pDR196 in yeast and they were transformed into the yeast mutant lack of oligopeptide transport function to test their oligopeptide transport function. [ Result] The actual and expected results of restriction enzyme digesting were same, indicating that the target genes had been cloned into the shuttle expression vector in yeast successfully. In the auxotrophic test on yeast strains, all the strains grew well in complete medium; the strains ABC154 and ABC738 could not grow in the media lack of Urn, Leu, Lys, His, Trp or Ade, indicating that the strains ABC154 and ABC738 were the auxotrophic strains of Urn, Leu, Lys, His, Trp and Ade and they could be used to test the oligopeptide transport function of target genes. The strains pDR196/OsPTR1 - 10/ ABC738 and pDR196/ABC738 grew well in the medium lack of Ura, indicating that there was Urn gene in the plasmid pDR196. [ Conclusion ] This experiment supplied methodological support for studying oligopeptide transport genes.