羊草钙依赖型蛋白激酶基因生物信息学及表达特性分析  

Analysis of Bioinformatics and Expression Pattern of Calcium-dependent Protein Kinase Gene in Leymus chinensis

作　　者：崔喜艳[1] 张继伟[2] 秦思余 刘忠野[1] 郭继勋[3] 

机构地区：[1]吉林农业大学生命科学学院,吉林长春130118 [2]东北师范大学附属中学,吉林长春130117 [3]东北师范大学草地研究所/植被生态科学教育部重点实验室,吉林长春130024

出　　处：《中国草地学报》2015年第3期11-18,共8页Chinese Journal of Grassland

基　　金：国家自然科学基金(30570273);吉林省教育厅项目(吉教科合字〔2009〕第61号)

摘　　要：为了研究钙依赖型蛋白激酶(CDPK)在羊草适应生境中的功能,分别对羊草幼苗进行模拟干旱(PEG)和盐碱(NaCl-Na2CO3)处理,测序分析了羊草中克隆的CDPK基因cDNA(LcCDPK),其核酸序列全长为1704bp,开放阅读框为1647bp,共编码548个氨基酸,编码的氨基酸序列与GenBank中小麦CDPK1基因的同源性最高,氨基酸一致性为96%。用软件对其蛋白质氨基酸组成及结构分析,氨基酸序列构成具有丝氨酸/苏氨酸蛋白激酶催化活性的S-TKc结构域,有4个保守的EF手性结构域,为Ca2+结合位点,预测的成熟肽相对分子质量61.349KDa,理论等电点5.34;二级结构预测α-螺旋占37.59%,β-折叠占9.85%,而无规则卷曲占52.55%;实时荧光定量PCR结果分析表明,在低浓度PEG、盐碱处理条件下羊草根及叶片的CDPK基因表达下调,高浓度处理条件下CDPK基因表达上调,说明CDPK基因表达受干旱及盐碱胁迫调控;盐碱处理条件下CDPK基因在根和叶片中的表达均高于干旱处理。推测LcCDPK广泛参与羊草抵御逆境胁迫的响应,尤其在盐碱胁迫过程中发挥重要作用。In order to study the function of calcium-dependent protein kinase （CDPK） in L. chinensis adapted to habitats, seedlings were treated respectively with simulation of drought （polyethylene glycol, PEG） and saline-alkali （NaC1-Na2 CO3）. This test sequenced and analyzed the cDNA of CDPK cloned from L. chinensis （LcCDPK）. The nucleic acid sequence length was 1704 bp, with an open reading frame （ORF） of 1647 bp, encoding 548 amino acids. The encoding amino acid sequence had the highest homology with CDPK1 of wheat from GenBank（96G）. The results of bioinformatics soft-wares showed： the a- mino acid sequence has a S-TKc structural domain with the catalytic activity of serine/threonine protein kinase, and there were four conservative EF-hands structural domain with binding Ca2＋. It was predicted that relative molecular mass of mature peptide was 61. 349 KDa and theoretical isoelectric point was 5.34, meanwhile, in secondary structure, alpha-helix accounted for 37. 59%, beta-fold 9. 85%, random coil 52.55%. The results of real-time PCR showed： CDPK gene expression in roots and leaves all were downregulating under low concentration PEG or saline alkali treatment conditions while its expression was up regulating under high concentration conditions. CDPK gene expression and regulation were impacted by drought and saline-alkali stress. CDPK gene expression with saline-alkali treatment all was higher than that of drought treatment in roots and leaves. Therefore, it is speculated that LcCDPK widely participate in defending stress in L. chinensis, and especially play an important role in the process of saline-alkali stress.

关 键 词：羊草 CDPK基因 生物信息学 基因表达 

分 类 号：Q943.2[生物学—植物学]