Li Duo-Chuan, Li Wei, Zhou Qing-Xin, Lu Jing, Peng You-Liang
Department of Environmental Biology, Shandong Agricultural University, Taian, Shandong, People's Republic of China.
DNA Seq. 2007 Dec;18(6):423-33. doi: 10.1080/10425170701318443.
Based on the conserved amino acid sequence (DLKPEN) of serine-threonine protein kinase from several fungi, a degenerate primer was designed and synthesized. Total RNA was isolated from the thermophilic fungus Thermomyces lanuginosus. Using RACE-PCR, full-length cDNA of a putative serine-threonine protein kinase gene was cloned from T. lanuginosus. The full-length cDNA of T. lanuginosus protein kinase was 2551 bp and contained an 1806 bp open reading frame encoding a putative protein kinase precursor of 601 amino acid residues. Sequencing analysis showed that the cloned cDNA of T. lanuginosus had consensus protein kinase sequences. Conservative amino acid subdomains which most serine-threonine kinases contain can be found in the deduced amino acid sequence of T. lanuginosus putative protein kinase. Comparison results showed that the deduced amino acid sequence of T. lanuginosus putative protein kinase was highly homologous to that of Neurospora crassa dis1-suppressing protein kinase Dsk1. The putative protein kinase contained three arginine/serine-rich (SR) regions and two transmembrane domains. These showed that it might be a novel putative serine-threonine protein kinase.
基于几种真菌丝氨酸 - 苏氨酸蛋白激酶的保守氨基酸序列（DLKPEN），设计并合成了简并引物。从嗜热真菌嗜热栖热菌中分离出总RNA。使用RACE-PCR从嗜热栖热菌中克隆了一个假定的丝氨酸 - 苏氨酸蛋白激酶基因的全长cDNA。嗜热栖热菌蛋白激酶的全长cDNA为2551 bp，包含一个1806 bp的开放阅读框，编码一个由601个氨基酸残基组成的假定蛋白激酶前体。测序分析表明，克隆的嗜热栖热菌cDNA具有一致的蛋白激酶序列。在嗜热栖热菌假定蛋白激酶的推导氨基酸序列中可以发现大多数丝氨酸 - 苏氨酸激酶所包含的保守氨基酸亚结构域。比较结果表明，嗜热栖热菌假定蛋白激酶的推导氨基酸序列与粗糙脉孢菌dis1抑制蛋白激酶Dsk1的序列高度同源。该假定蛋白激酶包含三个富含精氨酸/丝氨酸（SR）的区域和两个跨膜结构域。这些表明它可能是一种新型的假定丝氨酸 - 苏氨酸蛋白激酶。
