水稻OsHSP40基因可变剪接体鉴定及表达分析

Analyzing Splice Variants and Expression of the OsHSP40 Gene in Rice

综合运用生物信息学、RT-PCR和TA克隆技术对水稻热激蛋白基因OsHSP40的可变剪接体进行鉴定,并分析剪接体的表达。首先,利用生物信息学分析发现,该基因存在4种可变剪接体。根据不同可变剪接体的序列信息设计特异性引物进行RT-PCR扩增,结合TA克隆,成功鉴定到OsHSP40的4种可变剪接体。随后,提取野生型植株的根、茎、叶、幼穗和不同发育时期的籽粒,进行不同组织部位的表达分析,RT-PCR结果显示:可变剪接体4(OsHsp40.4)与OsHsp40.1234在不同组织部位的表达模式存在差异。最后,分析在高盐和高温条件下不同剪接本的表达,结果发现,高盐条件下OsHsp40.4和OsHsp40.1234表达上升,但OsHsp40.4上升幅度较低;高温处理后,OsHsp40.1234在处理后2 h表达变化不显著,处理后4 h其表达显著上升,而OsHsp40.4则在处理后2 h表达显著上升。综上,成功鉴定到OsHSP40基因的4种可变剪接体,发现可变剪接体OsHsp40.4和OsHsp40.1234在高盐和高温胁迫过程中的表达存在差异。上述结果为进一步研究该基因的生物学功能奠定基础。

To identify and analyze the expression of the splicers of OsHSP40 using bioinformatics,RT-PCR,TA cloning and sequencing.Firstly,there were 4 kinds of variable splices of OsHSP40 based on bioinformatics analysis.Specific primers were designed for RT-PCR amplification according to the sequence information of different variable splice variants,and combined with TA cloning,successfully identified four kinds of variable splice variants of OsHSP40.To investigate the expression pattern,total RNA was isolated from different tissues such as root,stem,leaf,panicle and seed of WT.RT-PCR results revealed that the expression patterns of OsHsp40.4 and OsHsp40.1234 were different.Finally,the expression of different splice variants under high salt and high temperature conditions was analyzed.The expressions of OsHsp40.4 and OsHsp40.1234 increased under high salt conditions,but the increase range of OsHsp40.4 was relatively low.In addition,after high temperature treatment,the expression of OsHsp40.1234 showed no significant difference at 2 h,and increased significantly at 4 h treatment,while the expression of OsHsp40.4 increased significantly at 2 h treatment.In conclusion,successfully identified four splice variants of OsHSP40,and found differences in the expression of OsHsp40.4 and OsHsp40.1234 in the process of high salt and high temperature stress.These results lay the foundation for further research on the biological function of this gene.