Knight J S, Gray J C
Department of Plant Sciences, University of Cambridge, UK.
Mol Gen Genet. 1994 Mar;242(5):586-94. doi: 10.1007/BF00285282.
A cDNA encoding the complete precursor of the phosphate translocator of the chloroplast inner envelope membrane has been isolated from a tobacco leaf (Nicotiana tabacum cv. Samsun) lambda gt 11 library. The tobacco cDNA is 1546 bp in length and encodes a precursor protein of 401 amino acid residues with a deduced molecular weight of 43705. A putative processing site between Ala-73 and Ala-74 of the precursor protein is suggested by comparison with the N-terminal sequences of the pea and spinach proteins. Removal of the transit peptide produces the mature protein of 328 amino acid residues with a molecular weight of 36038. Southern blot analysis suggests there is probably one copy of the phosphate translocator gene in the pea haploid genome and two copies in the tobacco haploid genome, one derived from each ancestral parental genome. Messenger RNAs essentially equivalent in size to the cDNAs (approx. 1.6 kb) were detected in extracts of all organs examined from tobacco and pea, including leaves, stems, sepals, petals, seed-pods, tendrils and roots. An immunochemically related protein of a similar size to the phosphate translocator was detected in the equivalent pea organs. The levels of both mRNA and protein in non-photosynthetic organs were lower than those in photosynthetic organs. Tobacco phosphate translocator mRNA was present at high levels in etiolated tissue and did not increase significantly after 24 h illumination. Germination and growth of tobacco seedlings in the presence of sucrose caused a 3.3-fold decrease in the level of the phosphate translocator mRNA.
已从烟草(Nicotiana tabacum cv. Samsun)λgt 11文库中分离出一个编码叶绿体内膜磷酸转运体完整前体的cDNA。烟草cDNA长度为1546 bp,编码一个由401个氨基酸残基组成的前体蛋白,推导分子量为43705。通过与豌豆和菠菜蛋白的N端序列比较,推测前体蛋白在Ala-73和Ala-74之间存在一个加工位点。去除转运肽后产生由328个氨基酸残基组成、分子量为36038的成熟蛋白。Southern杂交分析表明,豌豆单倍体基因组中可能有一个磷酸转运体基因拷贝,烟草单倍体基因组中有两个拷贝,分别来自每个祖先亲本基因组。在烟草和豌豆所有检测器官(包括叶、茎、萼片、花瓣、豆荚、卷须和根)的提取物中,检测到大小与cDNA基本相当(约1.6 kb)的信使RNA。在豌豆相应器官中检测到一种大小与磷酸转运体相似的免疫化学相关蛋白。非光合器官中mRNA和蛋白的水平均低于光合器官。烟草磷酸转运体mRNA在黄化组织中含量较高,光照24小时后无显著增加。在蔗糖存在下烟草幼苗的萌发和生长导致磷酸转运体mRNA水平下降3.3倍。