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以油果实为材料,应用RT-PCR和RACE技术克隆其β-tubulin cDNA,并采用半定量的方法对该基因进行表达量分析。结果表明,油果实β-tubulin的cDNA全长1606 bp,包含一个1341 bp的开放阅读框,共编码446个氨基酸。对其推导氨基酸序列进行比对分析,表明油果实β-tubulin与众多植物来源的β-tubulin具有很高的相似度。通过设计特异性引物,对果实11个发育时期β-tubulin表达量分析的结果表明,该β-tubulin基因表达量稳定,说明该β-tubulin可作为油果实整个生长发育时期内参基因使用,可以在为进一步应用实时荧光定量PCR技术研究油果实发育相关基因的表达奠定了试验基础。
The cDNA of β-tubulin was cloned by RT-PCR and RACE techniques. The expression of β-tubulin cDNA was analyzed by semi-quantitative method. The results showed that the β-tubulin cDNA was 1606 bp in length and contained a 1341 bp ORF encoding a total of 446 amino acids. The deduced amino acid sequences were compared and analyzed, which showed thatβ-tubulin in fruit tree had a high similarity withβ-tubulin derived from many plants. Through the design of specific primers, the results of β-tubulin expression in 11 developmental stages showed that the expression of β-tubulin gene was stable, which indicated that β-tubulin could be used as a reference gene for the whole growth and development stage of P. arvense. The experimental basis for further application of real-time fluorescence quantitative PCR technology to study the expression of fruit growth-related genes laid the foundation.