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【目的】克隆甘蔗MAP激酶家族新基因的全长序列,为了解甘蔗抗逆胁迫机制提供依据。【方法】以新台糖22为材料,提取甘蔗幼叶总RNA并反转录为cDNA;利用已知物种MAP激酶基因核苷酸序列保守区设计引物,采用5’和3’端RACE技术克隆新基因全长,并进行生物信息学分析。【结果】克隆获得的甘蔗新基因与玉米ZmMAPK4同源性很高,达92.6%,将该基因命名为SoMAPK4(登录号JQ062930),基因全长1499bp,其中开放阅读框(ORF)为1128bp,5’非翻译区(UTR)为218bp,3’非翻译区(UTR)为213bp。生物信息学分析结果表明,SoMAPK4基因编码一个含376个氨基酸的蛋白质,分子量约43.5kDa,等电点为5.51,含有11个保守的蛋白激酶亚区和MAP激酶的磷酸化位点TEY基序。【结论】克隆获得甘蔗MAP激酶新基因SoMAPK4,该基因可能参与多种胁迫反应的信号传递,是研究甘蔗非生物胁迫和生物胁迫过程中信号传递的一个关键因素。
【Objective】 Cloning the full-length sequence of the new gene of the MAP kinase family in sugarcane provides a basis for understanding the mechanism of stress tolerance in sugarcane. 【Method】 The total RNA of young leaves of sugarcane was extracted with Xintai22 and the total RNA was reverse transcribed into cDNA. The primers were designed based on the conserved region of the nucleotide sequence of MAP kinase gene of known species. The 5 ’and 3’ Gene length, and bioinformatics analysis. 【Result】 The results showed that the new gene of cloned sugarcane had high homology with ZmMAPK4 in maize, accounting for 92.6%. The gene was named SoMAPK4 (accession number JQ062930) and the full-length gene was 1499bp. The open reading frame (ORF) was 1128bp, 5 The untranslated region (UTR) was 218 bp and the 3 ’untranslated region (UTR) was 213 bp. The results of bioinformatics analysis showed that SoMAPK4 gene encodes a 376 amino acid protein with a molecular weight of about 43.5 kDa and an isoelectric point of 5.51 and contains 11 conserved protein kinase subtypes and MAP kinase phosphorylation site TEY motifs. 【Conclusion】 SoMAPK4, a new gene of sugarcane MAP kinase, was cloned and may be involved in the signal transduction of many stress responses. It is a key factor to study the signal transduction in sugarcane abiotic stress and biotic stress.