铁皮石斛可溶性酸性转化酶基因克隆与表达分析

发布时间：2018-03-07 09:12

本文选题：铁皮石斛　切入点：酸性转化酶　出处：《华南农业大学学报》2017年02期 　论文类型：期刊论文

【摘要】：【目的】克隆铁皮石斛Dendrobium officinale可溶性酸性转化酶基因SAI,分析该基因生物学信息及时空表达特异性。【方法】基于同源序列克隆铁皮石斛SAI基因c DNA全长,采用生物信息学方法进行序列分析,并采用qRTPCR进行组织特异性表达分析。【结果】铁皮石斛SAI基因全长为1 595 bp,c DNA编码区1 368 bp,Genbank登录号为KU598852。该基因编码455个氨基酸,蛋白相对分子质量为50 700。NCBI BLASTx分析表明,该基因氨基酸序列与柑橘Citrus unshiu酸性转化酶基因、甜橙C.sinensis酸性β-呋喃果糖苷酶基因序列相似性最高,达77%,与其他植物酸性转化酶基因的相似性均高于68%。聚类分析表明,铁皮石斛SAI基因与玉米Zea mays液泡转化酶基因、甘蔗Saccharum officinarum SAI基因亲缘关系最近。该基因编码的蛋白质属于非跨膜结构的亲水性热稳定蛋白,定位于液泡中。SAI基因在2年生铁皮石斛的茎中表达量最高,3年生的叶中表达量最低。【结论】成功克隆铁皮石斛液泡酸性转化酶基因,该基因在铁皮石斛不同组织不同生育时期的表达量差异较大。
[Abstract]:[objective] to clone the soluble acid invertase gene of Dendrobium officinale from Dendrobium candidum and analyze its biological information and spatio-temporal expression specificity. [methods] based on the homologous sequence, the full length of c DNA of SAI gene of Dendrobium candidum was cloned. Sequence analysis was carried out by bioinformatics, and tissue specific expression was analyzed by qRTPCR. [results] the total length of SAI gene of Dendrobium candidum was 1 595 BP C DNA coding region 1 368 BP Genbank accession number was KU598852. The gene encodes 455 amino acids. The protein molecular weight was 50,700.NCBI BLASTx analysis showed that the amino acid sequence of this gene was the most similar to that of Citrus unshiu acid invertase gene and C. sinensis acid 尾 -furan fructosidase gene. The similarity with other plant acid invertase genes was higher than 68. Cluster analysis showed that the SAI gene of Dendrobium candidum and the Zea mays vacuolar invertase gene of maize were higher than those of other plants. The Saccharum officinarum SAI gene of sugarcane is closely related. The protein encoded by this gene belongs to the hydrophilic thermostable protein with non-transmembrane structure. The expression of SAI gene was the highest in the stems of 2-year-old Dendrobium candidum and the lowest in the leaves of 3-year old Dendrobium candidum. [conclusion] the vacuolar acid invertase gene of Dendrobium candidum was cloned successfully. The expression of this gene in different tissues of Dendrobium candidum was different in different growth stages.
【作者单位】：云南省高校农作物优质高效栽培与安全控制重点实验室/红河学院生命科学与技术学院;
【分类号】：S828

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