Hayashida N, Mizoguchi T, Yamaguchi-Shinozaki K, Shinozaki K
Laboratory of Plant Molecular Biology, Tsukuba Life Science Center, Ibaraki, Japan.
Gene. 1992 Nov 16;121(2):325-30. doi: 10.1016/0378-1119(92)90138-f.
Cloning and analysis of a gene that encodes a homologue of protein kinase (PK) from Arabidopsis thaliana is reported. Oligodeoxyribonucleotides (oligos) corresponding to conserved regions in catalytic domains of various PKs were used for polymerase chain reaction (PCR) amplification with genomic DNA from A. thaliana as template, in an attempt to identify genes encoding PK in plants. We obtained several amplified DNA fragments that encoded part of a PK. We screened a genomic DNA library of A. thaliana with these oligos or PCR fragments as probes. Three genomic clones were obtained and one of them, named Atpk7, was sequenced and analyzed. Atpk7 was demonstrated by PCR to contain an intron. The mRNA transcribed from Atpk7 was detected predominantly in root tissue by Northern blot analysis. The transcription start point was determined by primer extension. The deduced amino acid (aa) sequence of the putative product of Atpk7 resembles those of S6 kinases, cyclic nucleotide-dependent PKs and calcium-dependent PKs. From this comparison of aa sequences, the ATPK7 protein is considered to be a member of a novel subfamily of Ser/Thr PKs in plants.
报道了拟南芥中一个编码蛋白激酶（PK）同源物的基因的克隆与分析。以拟南芥基因组DNA为模板，使用与各种PK催化结构域保守区域相对应的寡脱氧核糖核苷酸（oligos）进行聚合酶链反应（PCR）扩增，试图鉴定植物中编码PK的基因。我们获得了几个编码PK部分序列的扩增DNA片段。我们用这些oligos或PCR片段作为探针筛选了拟南芥基因组DNA文库。获得了三个基因组克隆，其中一个命名为Atpk7，进行了测序和分析。通过PCR证明Atpk7含有一个内含子。通过Northern印迹分析在根组织中主要检测到从Atpk7转录的mRNA。通过引物延伸确定转录起始点。Atpk7推定产物的推导氨基酸（aa）序列与S6激酶、环核苷酸依赖性PK和钙依赖性PK的序列相似。通过氨基酸序列的比较，认为ATPK7蛋白是植物中Ser/Thr PK新亚家族的成员。
