We isolated the rye serine/threonine protein kinase (S/T PK) gene, which is responsible for intracellular signal transduction mediated by abscisic acid (ABA) synthesized following drought stress, from the huge genome, i.e., 1Cx = 7917 Mb. cDNA was synthesized from poly(A)+ RNA from ABA-treated leaves and then ligated with pUC118 via EcoRI linker to form a cDNA library. RT-PCR amplified DNA fragment of S/T PK gene was used as a probe to screen the cDNA library. One positive cDNA clone was sequenced to determine the overall 1462 bp of the S/T PK gene, which encodes the S/T PK protein consisting of deduced 363 amino acid residues including 12 catalytic subdomains. Total RNA was extracted from both ABA-treated and ABA-untreated rye Petkus leaves and then run on a 1% agarose gel denatured by formaldehyde, and the RNA was transblotted to a nylon membrane. Next, northern hybridization was conducted on the membrane using the 3′ untranslated 244 bp region as probes. The rye S/T PK gene was transcribed in ABA-treated leaves but not in untreated leaves. We have shown that rye S/T PK, localized on the 2R chromosome, is transcribed via ABA mediation, which indicates the relation to intracellular signal transduction.

我们从庞大的基因组（即1Cx = 7917 Mb）中分离了黑麦丝氨酸/苏氨酸蛋白激酶（S / T PK）基因，该基因负责干旱胁迫后合成的脱落酸（ABA）介导的细胞内信号转导。从ABA处理的叶片的poly（A）+ RNA合成cDNA，然后通过Eco将其与pUC118连接RI接头形成cDNA文库。用RT-PCR扩增的S / T PK基因DNA片段作为探针筛选cDNA文库。对一个阳性cDNA克隆进行测序以确定S / T PK基因的总1462 bp，该基因编码S / T PK蛋白，该蛋白由推导的363个氨基酸残基组成，包括12个催化亚结构域。从经过ABA处理和未经ABA处理的黑麦Petkus叶片中提取总RNA，然后在经过甲醛变性的1％琼脂糖凝胶上电泳，并将RNA转移到尼龙膜上。接下来，使用3'非翻译的244bp区域作为探针在膜上进行RNA杂交。黑麦S / T PK基因在ABA处理的叶片中转录，但未在未处理的叶片中转录。我们已经证明，位于2R染色体上的黑麦S / T PK是通过ABA介导转录的，