Schultz S J, Nigg E A
Swiss Institute for Experimental Cancer Research, Epalinges.
Cell Growth Differ. 1993 Oct;4(10):821-30.
The nimA gene encodes a protein-serine/threonine kinase that is required along with the p34cdc2 kinase for mitosis in Aspergillus nidulans. We have searched for human protein kinases that are related to the NIMA protein kinase using the polymerase chain reaction. Different pairs of degenerate oligonucleotides specific for conserved amino acid motifs in the catalytic domain of NIMA were used as primers in the polymerase chain reaction to amplify partial complementary DNAs (cDNAs) of protein kinases expressed in the promyelocytic leukemia cell line HL-60. Forty-one distinct cDNAs representing a broad spectrum of serine/threonine- and tyrosine-specific protein kinases were identified, and the sequences for 21 of these protein kinases were found to be unique. Three of these cDNAs represent a family of protein kinases whose members are related to NIMA and the murine nimA-related protein kinase Nek1. We discuss the success of this polymerase chain reaction approach with respect to the use of multiple primer pairs, the influence of primer degeneracy, and the tolerance of cDNA amplification to mismatches between primers and template mRNA.
nimA基因编码一种蛋白质丝氨酸/苏氨酸激酶，在构巢曲霉中，它与p34cdc2激酶一起是有丝分裂所必需的。我们利用聚合酶链反应寻找与NIMA蛋白激酶相关的人类蛋白激酶。针对NIMA催化结构域中保守氨基酸基序的不同简并寡核苷酸对被用作聚合酶链反应的引物，以扩增早幼粒细胞白血病细胞系HL-60中表达的蛋白激酶的部分互补DNA（cDNA）。鉴定出41个代表广泛的丝氨酸/苏氨酸和酪氨酸特异性蛋白激酶的不同cDNA，其中21个蛋白激酶的序列是独特的。这些cDNA中的三个代表一个蛋白激酶家族，其成员与NIMA和小鼠NIMA相关蛋白激酶Nek1有关。我们讨论了这种聚合酶链反应方法在使用多个引物对方面的成功、引物简并性的影响以及cDNA扩增对引物与模板mRNA之间错配的耐受性。
