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细胞色素P450基因在电子传递链、次生代谢物质合成和对外源化学药物毒性降解中发挥着重要作用,为了深入了解该基因在甘蔗中的功能,通过RT-PCR扩增获得甘蔗细胞色素P450还原酶基因的cDNA全长序列,命名为ScCPR450(Gen Bank Accession Number:KR864841).该基因全长999 bp,含有744 bp的完整开放阅读框,编码247个氨基酸.亚细胞定位结果显示,ScCPR450蛋白分布于细胞质中,与生物信息学预测结果相符.q RT-PCR表达分析表明,该基因在甘蔗中组成型表达,但有组织特异性,芽中表达量最高,其次是叶,而皮中表达量最低.在脱落酸(ABA)、水杨酸(SA)、茉莉酸甲酯(Me JA)、聚乙二醇(PEG)和氯化铜(CuCl_2)胁迫诱导过程中,该基因的表达量呈现不同变化模式,其中SA胁迫6 h下,ScCPR450基因的表达量最高,约为对照的12.21倍;在PEG胁迫下,ScCPR450基因的表达量上调且表达量稳定,推测ScCPR450基因在甘蔗响应生物和非生物胁迫中发挥一定的作用.本研究可为该基因家族其它成员的克隆以及深入解析该基因的功能特性奠定基础,进而为基于基因工程技术对甘蔗品种进行定向改良提供基因资源.
Cytochrome P450 plays an important role in the electron transfer chain, the synthesis of secondary metabolites and the toxicity degradation of exogenous chemical drugs. To understand the function of this gene in sugarcane, the cytochrome P450 gene is amplified by RT-PCR The complete cDNA sequence of the gene was designated as ScCPR450 (GenBank Accession Number: KR864841). The full-length cDNA was 999 bp in length and contained a 744 bp open reading frame encoding a protein of 247 amino acids.The subcellular localization of ScCPR450 protein In the cytoplasm, it was consistent with the prediction of bioinformatics.q-RT-PCR analysis showed that the gene was constitutively expressed in sugarcane but tissue-specific, with highest expression in shoots, followed by leaves, while expression in skin The lowest was in the course of ABA, SA, Me JA, PEG and CuCl 2 stress induction Under different stress modes, the expression level of ScCPR450 gene was the highest at 6 h after SA stress, which was about 12.21 times of that of control. Under the stress of PEG, the expression level of ScCPR450 gene was up-regulated and the expression level was stable. Material stress plays a role in this research cloning other members of the gene family can lay the foundation for in-depth analysis and functional properties of the gene, thus providing genetic resources to be directed to improvement of sugarcane varieties based on genetic engineering techniques.