根据小麦CIPK相关EST序列TC95390,采用eDNA末端快速扩增(RACE)技术,从耐盐性、耐旱性较好的小麦品种石4185中克隆出5’末端和3’末端,序列拼接后获得全长基因,同源比对后命名为TaClPK31(Gen-Bank登录号JX625142.1),并对该基因进行了生物信息学分析.结果表明,TaCIPK31编码区长1350bp,编码449个氨基酸;TaCIPK31的理论等电点为8.27,相对分子质量为50.9kD,是亲水性较强的氨基酸;TaCIPK31的N-端激酶催化域含有该家族基因的典型激活环,C-端调节域含有CIPK家族中高度保守的NAF结构域,具有丝氨酸-苏氨酸蛋白激酶的结构特征;氨基酸序列同源性分析表明,TaCIPK31与已报道的大麦HvCIPK31、水稻OsCIPK3、玉米ZmCIPK31有较高的同源性.
Based on the wheat CIPK EST sequence of TC95390, we got the 3'-end and 5'-end using RACE (rapid-amplification of cDNA ends) method from wheat cuhivar Shi 4185 which has higher drought-resistance and salt-resistance. The whole sequence of gene was analyzed and named as TaCIPK31 by sequence homology and deposited in GenBank database of accession number JX625142.1. Bioinformatics analysis was undergone meanwhile. The results showed that the ORF length of TaCIPK31 was 1 350 bp and encoded 449 amino acids. The pI and Mw of TaCIPK31 were 8.27 and 50.9 kD, respectively. TaCIPK31 was a hydrophilic stable protein while there was only one significant hydrophobic peak and one transmembrane peptide. The TaCIPK31 protein contained a catalytic Ser/ Thr protein domain in its N-terminal region and a regulatory domain with NAF motif at its C-terminus, which is in line with the typical CIPK structural characteristics. The amino acids sequence analysis showed that TaCIPK31 has higher homology with HvCIPK31, OsCIPK3 and ZmCIPK31.