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可变剪接(Alternative splic ing,AS)是真核生物基因表达调控研究的热点.BAK1(Brassinosteroid insensitive1-associated receptor kinase 1)是植物中丝氨酸/苏氨酸蛋白激酶的一种,可调控植物的生长发育及先天免疫反应.为揭示BAK1基因在甘蔗应答不良外界环境方面的作用,利用电子克隆及RT-PCR方法从甘蔗品种崖城05-179叶片c DNA中克隆到1个BAK1基因及其1个可变剪接体,分别命名为Sc BAK1(Gen Bank登录号:KP032226)和Sc BAK1 S1(Gen Bank登录号:KP032227).生物信息学分析结果表明,Sc BAK1/Sc BAK1 S1基因的ORF长1 860 bp/1 770 bp,编码蛋白含有619/589个氨基酸残基,分子量(Mr)为6.928×104/6.576×104;两种编码蛋白均定位于质膜上,含有信号肽,为酸性、分泌脂溶性蛋白;它们的二级结构以α-螺旋和无规则卷曲为主,其次是延伸链,无β-螺旋;蛋白功能预测显示其主要作为细胞膜蛋白,还参与氨基酸及辅酶因子生物合成.荧光定量PCR分析结果显示,Sc BAK1 S1基因的表达在非生物(水杨酸SA、Cu Cl2、PEG、脱落酸ABA、Na Cl、茉莉酸甲酯JA)和生物胁迫(黑穗病菌)下均受到抑制,而Sc BAK1却受SA、Cu Cl2、PEG、Na Cl和黑穗病菌的诱导.结果还表明,相较于Sc BAK1在甘蔗抗黑穗病性、渗透胁迫以及细胞生长方面发挥作用来说,Sc BAK1 S1缺失的氨基酸序列或数目在Sc BAK1的抗逆性方面扮演了重要角色.Sc BAK1和Sc BAK1 S1的不同丰度表达为深入解析Sc BAK1基因在生物和非生物逆境条件下的功能奠定了基础.
Alternative splicing (AS) is a hotspot in the regulation of gene expression in eukaryotes.BAK1 (Brassinosteroid insensitive1-associated receptor kinase 1) is a kind of serine / threonine protein kinase in plants that regulates plant growth Development and innate immunity.In order to reveal the role of BAK1 gene in the poor external environment of sugarcane response, one BAK1 gene and one BAK1 gene were cloned from the c DNA of Yacheng 05-179 cane by using electronic cloning and RT-PCR. ScBAK1 (GenBank accession number: KP032226) and Sc BAK1 S1 (GenBank accession number: KP032227), respectively.The bioinformatics analysis showed that the ORF of Sc BAK1 / Sc BAK1 S1 gene was 1860 bp / 1 770 bp. The encoded protein contained 619/589 amino acid residues and the molecular weight (Mr) was 6.928 × 104 / 6.576 × 104. Both encoded proteins were located on the plasma membrane and contained signal peptide, which was acidic and secreted lipid Soluble protein.These secondary structures were mainlyα-helix and random coil, followed by extension chain withoutβ-helix.The prediction of protein function showed that it was mainly involved in cell membrane protein and also participated in the biosynthesis of amino acids and coenzyme.Fluorescence quantification PC R analysis showed that the expression of Sc BAK1 S1 gene was inhibited under abiotic conditions (SA, Cu Cl2, PEG, abscisic acid ABA, Na Cl, JA) and biotic stress , While Sc BAK1 was induced by SA, Cu Cl2, PEG, Na Cl and S. gossypii.The results also showed that compared to Sc BAK1, which plays a role in anti-smut resistance, osmotic stress and cell growth in sugarcane, The amino acid sequence or number of deletion of Sc BAK1 S1 plays an important role in the resistance of Sc BAK1 The different abundance of Sc BAK1 and Sc BAK1 S1 is expressed as an insight into the function of the Sc BAK1 gene under both biological and abiotic stress conditions The foundation.