Xu Y, Buchholz W G, DeRose R T, Hall T C
Institute of Developmental and Molecular Biology, Texas A&M University, College Station 77843-3155.
Plant Mol Biol. 1995 Jan;27(2):237-48. doi: 10.1007/BF00020180.
Two cDNA clones (RCc2 and RCc3) corresponding to mRNAs highly expressed only in root tissues of rice (Oryza sativa L.) seedlings were characterized. Respectively, they encode polypeptides of 146 (14.5 kDa) and 133 amino acids (13.4 kDa) that share high (> 70%) sequence similarity with a polypeptide encoded by a cDNA (ZRP3) encoding an mRNA preferentially expressed in young maize roots. Genomic DNA blot analysis revealed that they are members of a small gene family and RCg2, the gene corresponding to RCc2, was isolated. A 1656 bp 5'-upstream sequence of RCg2 was translationally fused to a beta-glucuronidase (GUS) reporter gene and stable introduction of the chimeric construct into rice was confirmed by PCR and genomic DNA blot analyses. Histochemical analysis of transgenic rice plants containing the full-length chimeric gene showed high levels of GUS activity in mature cells and the elongation and maturation zones of primary and secondary roots, and in the root caps, but no GUS activity was detected in root meristematic regions. Surprisingly, high GUS activity was also detected in leaves of the same plants. This raises the possibility that the RCg2 5'-upstream element may not be sufficient for the proper spatial control of root specificity in transgenic rice.
对两个与仅在水稻(Oryza sativa L.)幼苗根组织中高表达的mRNA相对应的cDNA克隆(RCc2和RCc3)进行了表征。它们分别编码146个氨基酸(14.5 kDa)和133个氨基酸(13.4 kDa)的多肽,这些多肽与由优先在玉米幼根中表达的mRNA的cDNA(ZRP3)编码的多肽具有高度(> 70%)的序列相似性。基因组DNA印迹分析表明它们是一个小基因家族的成员,并分离出了与RCc2对应的基因RCg2。将RCg2的1656 bp 5'-上游序列与β-葡萄糖醛酸酶(GUS)报告基因进行翻译融合,并通过PCR和基因组DNA印迹分析证实了嵌合构建体在水稻中的稳定导入。对含有全长嵌合基因的转基因水稻植株的组织化学分析表明,在成熟细胞、初生根和次生根的伸长区和成熟区以及根冠中检测到高水平的GUS活性,但在根分生组织区域未检测到GUS活性。令人惊讶的是,在同一植株的叶片中也检测到了高GUS活性。这增加了RCg2 5'-上游元件可能不足以在转基因水稻中对根特异性进行适当空间控制的可能性。