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为了解剪接因子在棘腹蛙发育过程中的功能,首先基于棘腹蛙转录组数据库的组装结果设计特异性引物,利用RT-PCR方法克隆到棘腹蛙体内编码丝氨酸/精氨酸富集剪接因子2(Serine/Arginine-rich splicing factor 2,SRSF2)的完整开放读码框(ORF),其ORF长度为642 bp,编码213个氨基酸残基,利用数字表达谱和半定量RT-PCR的方法验证了SRSF2在不同温度下的表达图谱,对克隆到的SRSF2基因进行了内含子分析。结果表明:SRSF2在不同生长温度(15,21,27,30℃)下的表达量存在显著差异,在21℃环境下的表达量最高,而在15,27,30℃生长环境下的表达量都较低。说明棘腹蛙来源的SRSF2基因ORF序列内包含个1内含子和2个外显子。
In order to understand the function of splicing factors during the development of Spinacia frugiperda, specific primers were designed based on the assembly results of Spinobagrus palustris transcriptome database and cloned into Serine / Arginine-rich splicing in vivo by RT-PCR The complete open reading frame (ORF) of Serine / Arginine-rich splicing factor 2 (SRSF2), whose ORF length is 642 bp and encodes 213 amino acid residues, was analyzed by using digital expression profiling and semi-quantitative RT-PCR The expression pattern of SRSF2 at different temperatures was verified, and the intron analysis of the cloned SRSF2 gene was performed. The results showed that there was significant difference in the expression of SRSF2 at different growth temperature (15, 21, 27, 30 ℃), and highest expression at 21 ℃, while the expression of SRSF2 at 15, 27, 30 ℃ Are lower. These results indicated that the ORF sequence of SRSF2 gene derived from Sparrow frog contains one intron and two exons.