青海茄参McCCoAOMT基因的克隆及其在冷胁迫下的表达分析  

McCCoAOMT gene cloning in Mandragora chinghaiensis and its expression analysis under cold stress

作　　者：王玉洁 宋帅 赵艳艳 WANG Yujie;SONG Shuai;ZHAO Yanyan(College of Agriculture and Animal Husbadry,Qinghai University,Xining 810016,China)

机构地区：[1]青海大学农牧学院,西宁810016

出　　处：《青海大学学报》2025年第1期55-61,共7页Journal of Qinghai University

基　　金：国家自然科学基金(31860575)。

摘　　要：为探究冷胁迫条件下青海茄参McCCoAOMT基因的调控机理,利用RT-PCR技术克隆该基因,并对其进行生物信息学及冷胁迫表达分析。结果表明:(1)McCCoAOMT为典型的腺苷三磷酸转移酶,是AdoMet-MTases基因家族中的一员。青海茄参McCCoAOMT基因的开放阅读框(ORF)长度为216 bp,编码71个氨基酸。(2)青海茄参与其他12个物种的CCoAOMT蛋白均包含保守的SAM结构域。青海茄参McCCoAOMT蛋白与番茄(NP_001234801.1)CCoAOMT蛋白的亲缘关系最近。(3)青海茄参McCCoAOMT蛋白的分子式为C_(340)H_(556)N_(90)O_(106)S_(6),分子量为7793.05 Da,理论等电点为8.03(>7);McCCoAOMT为稳定亲水性蛋白,其亚细胞定位于细胞质中,以无规则卷曲与α-螺旋作为二三级的主要结构;McCCoAOMT蛋白无跨膜结构,无信号肽,含有5个丝氨酸、2个苏氨酸和0个酪氨酸位点。(4)在冷胁迫条件下,青海茄参叶片中的McCCoAOMT基因表达量呈先上升后下降再上升的趋势,在处理24 h时其相对表达量达到最高。研究结果可为进一步探究青海茄参的耐寒分子机制提供一定的数据参考。To investigate the regulatory mechanism of McCCoAOMT gene in Mandragora chinghaiensis under cold stress,the gene was cloned using RT-PCR technology to perform bioinformatics and cold stress expression analysis.The results are as follows:(1)McCCoAOMT was a typical adenosine triphosphate transferase,which was a member of AdoMet-MTases gene family.The open reading frame(ORF)of McCCoAOMT gene of M.chinghaiensis was 216 bp in length,encoding 71 amino acids.(2)The CCoAOMT proteins of M.chinghaiensis and 12 other species all contain conserved SAM domains.The genetic relationship between McCCoAOMT protein of M.chinghaiensis and CCoAOMT protein of Solanum lycopersicum(NP_001234801.1)was the closest.(3)The molecular formula of McCCoAOMT protein in M.chinghaiensis was C_(340)H_(556)N_(90)O_(106)S_(6),with a molecular weight of 7793.05 Da and a theoretical isoelectric point of 8.03(>7);McCCoAOMT was a stable hydrophilic protein that was subcellular localized in the cytoplasm,with irregular curls andα-helices as main secondary and tertiary structures;McCCoAOMT protein contained 5 serine,2 threonine and 0 tyrosine sites,without trans membrane structure and signal peptide.(4)Under cold stress,the expression level of McCCoAOMT gene in the leaves of M.chinghaiensis showed a trend of first increasing,then decreasing and then increasing again,and its relative expression level reached the highest at 24 h of treatment.The research results can provide data reference for further exploring the molecular mechanism of cold resistance in Mandragora chinghaiensis.

关 键 词：青海茄参 基因克隆 表达分析 咖啡酰辅酶A-O-甲基转移酶 

分 类 号：Q789[生物学—分子生物学] S641.9[农业科学—蔬菜学]