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本研究利用Genefishing技术经差异表达技术从花生冷胁迫中分离生长素抑制蛋白基因,经荧光定量PCR验证,随着种子冷胁迫处理时间的延长,该基因在耐冷种子中表达量升高。经RACE技术得到该基因的c DNA全长序列,经序列拼接后,获得738 bp全长基因,开放式阅读框为342 bp,编码113个氨基酸,推测分子量为12 282.7 Da,理论等电点为9.81。信号肽预测不属于分泌蛋白。多重序列比对结果表明,其具有生长素抑制基因家族的保守结构域。本研究为该基因在花生耐冷性转基因功能分析提供依据。
In this study, Genefishing was used to isolate the auxin gene from cold stress of peanut by differential expression technique. The results showed that the expression level of the gene in cold-tolerant seed increased with the prolonging of cold stress treatment. The full-length cDNA of cDNA was obtained by RACE. The full-length cDNA of 738 bp was obtained by sequencing. The open reading frame was 342 bp, encoding 113 amino acids. The deduced molecular weight was 12 282.7 Da. The theoretical isoelectric point 9.81. Signal peptide is not predicted to be secreted protein. Multiple sequence alignment showed that it has a conserved domain of the auxin repressor gene family. This study provides a basis for the analysis of gene function of cold tolerance in peanut.