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拟南芥第二个肽转运基因的克隆

Cloning of a second Arabidopsis peptide transport gene.

作者信息

Song W, Steiner H Y, Zhang L, Naider F, Stacey G, Becker J M

机构信息

Department of Microbiology, University of Tennessee, Knoxville 37996-0845, USA.

出版信息

Plant Physiol. 1996 Jan;110(1):171-8. doi: 10.1104/pp.110.1.171.

DOI:10.1104/pp.110.1.171
PMID:8587981
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC157706/
Abstract

Previously, we reported the isolation of a peptide transport gene designated AtPTR2 from Arabidopsis thaliana by functional complementation of a yeast peptide transport mutant. We now report the isolation of a second peptide transport gene (AtPTR2-B) from Arabidopsis using the same approach. Similar to the effects of transferring AtPTR2-A (previously called AtPTR2), transfer of AtPTR2-B to yeast peptide transport mutants restored the ability to grow on di- and tripeptides but not peptides four residues or longer. However, unlike yeast mutants complemented with either the yeast PTR2 gene or the AtPTR2-A gene, transformants expressing AtPTR2-B were only partially sensitive to toxic peptides. Northern analysis showed that AtPTR2-B was constitutively expressed in all plant organs. Studies of the kinetics indicated that AtPTR2-A and AtPTR2-B have Km values of 47 and 14 microM, respectively, with Vmax values of 0.061 and 0.013 nmol mg-1 cell dry weight s-1, respectively, when dileucine was used as a substrate. AtPTR2-B is encoded on a 2.0-kb cDNA corresponding to a 585-amino acid protein (64.4 kD). Hydropathy analysis indicates that the protein is highly hydrophobic and suggests that there are 12 putative transmembrane segments. AtPTR2-B, like AtPTR2-A, shares significant similarity to a number of other proteins involved in transport of peptides into cells.

摘要

此前,我们报道了通过酵母肽转运突变体的功能互补从拟南芥中分离出一个名为AtPTR2的肽转运基因。我们现在报告使用相同方法从拟南芥中分离出第二个肽转运基因(AtPTR2-B)。与转移AtPTR2-A(先前称为AtPTR2)的效果类似,将AtPTR2-B转移到酵母肽转运突变体中恢复了在二肽和三肽上生长的能力,但不能在四个残基或更长的肽上生长。然而,与用酵母PTR2基因或AtPTR2-A基因互补的酵母突变体不同,表达AtPTR2-B的转化体仅对有毒肽部分敏感。Northern分析表明AtPTR2-B在所有植物器官中组成性表达。动力学研究表明,当以双亮氨酸作为底物时,AtPTR2-A和AtPTR2-B的Km值分别为47和14 microM,Vmax值分别为0.061和0.013 nmol mg-1细胞干重s-1。AtPTR2-B由一个2.0-kb的cDNA编码,对应于一个585个氨基酸的蛋白质(64.4 kD)。亲水性分析表明该蛋白质具有高度疏水性,并表明有12个推定的跨膜区段。AtPTR2-B与AtPTR2-A一样,与许多其他参与将肽转运到细胞中的蛋白质具有显著相似性。

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