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OALib Journal期刊
ISSN: 2333-9721
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甘蔗scbak1基因及其可变剪接体的克隆与表达分析
DOI: 10.3724/SP.J.1145.2015.03005, PP. 872-881
Keywords: 甘蔗,bak1基因,可变剪接,生物信息学,荧光定量pcr
Abstract:
可变剪接（alternativesplicing，as）是真核生物基因表达调控研究的热点.bak1（brassinosteroidinsensitive1-associatedreceptorkinase1）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cdna中克隆到1个bak1基因及其1个可变剪接体，分别命名为scbak1（genbank登录号：kp032226）和scbak1s1（genbank登录号：kp032227）.生物信息学分析结果表明，scbak1/scbak1s1基因的orf长1860bp/1770bp，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，scbak1s1基因的表达在非生物（水杨酸sa、cucl2、peg、脱落酸aba、nacl、茉莉酸甲酯ja）和生物胁迫（黑穗病菌）下均受到抑制，而scbak1却受sa、cucl2、peg、nacl和黑穗病菌的诱导.结果还表明，相较于scbak1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，scbak1s1缺失的氨基酸序列或数目在scbak1的抗逆性方面扮演了重要角色.scbak1和scbak1s1的不同丰度表达为深入解析scbak1基因在生物和非生物逆境条件下的功能奠定了基础.
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