Bartholomew D M, Rees D J, Rambaut A, Smith J A
Department of Plant Sciences, University of Oxford, UK.
Plant Mol Biol. 1996 May;31(2):435-42. doi: 10.1007/BF00021806.
We report the sequence of a cDNA clone encoding the c ("16 kDa') subunit of a vacuolar-type H(+)-ATPase (V-ATPase) from Kalanchoë daigremontiana, a plant in which the cell vacuole plays a pivotal role in crassulacean acid metabolism. The clone, pKVA211, was isolated from a K. daigremontiana leaf cDNA library constructed in lambda ZAP II using a homologous PCR-generated cDNA probe for the V-ATPase c subunit. The KVA211 cDNA was 839 nucleotides long and included a 20 bp poly(A)+ tail together with a complete 495 bp coding region for a polypeptide with a predicted molecular mass of 16659 Da. The deduced amino acid sequence was highly conserved across the wide range of eukaryotes (vertebrates, invertebrates, fungi, plants and protozoa) in which this gene has now been identified. Sequence comparison of several PCR products and genomic Southern analysis indicated that the V-ATPase c subunit in K. daigremontiana is encoded by a small multi-gene family. Steady-state levels of the KVA211 mRNA were much higher in leaves than in roots or flowers, and expression of this transcript in leaves was shown to be strongly light-dependent.
我们报道了编码来自落地生根（一种其细胞液泡在景天酸代谢中起关键作用的植物）的液泡型H(+)-ATP酶（V-ATP酶）的c（“16 kDa”）亚基的cDNA克隆序列。该克隆pKVA211是使用针对V-ATP酶c亚基的同源PCR产生的cDNA探针，从构建于λZAP II中的落地生根叶片cDNA文库中分离得到的。KVA211 cDNA长839个核苷酸，包括一个20 bp的聚腺苷酸尾巴以及一个完整的495 bp编码区，编码一个预测分子量为16659 Da的多肽。推导的氨基酸序列在现已鉴定出该基因的广泛真核生物（脊椎动物、无脊椎动物、真菌、植物和原生动物）中高度保守。几种PCR产物的序列比较和基因组Southern分析表明，落地生根中的V-ATP酶c亚基由一个小的多基因家族编码。KVA211 mRNA的稳态水平在叶片中比在根或花中高得多，并且该转录本在叶片中的表达显示出强烈的光依赖性。
