扁穗牛鞭草actin基因克隆及荧光定量PCR的建立

Clone actin gene and estiablishment of quantitative real-time PCR from Hemarthria compressa

搜索GenBank中禾本科植物actin基因序列进行同源性比对,选择保守区段设计简并引物,利用RT-PCR方法从扁穗牛鞭草中克隆获得actin基因cDNA片段。该基因片段长度为773 bp,编码257个氨基酸残基。根据此actin基因序列设计引物,建立了基于SYBR Green I染料技术的实时荧光定量PCR方法。该方法扩增效率高,检测范围广(R2=0.995,E=101.3%),为actin基因作为内参基因进行扁穗牛鞭草功能基因的定量分析奠定了基础。

Through homology comparison of actin gene sequences of gramineous plants from GenBank, conservative regions were selected for the designation of primers. The partial cDNA fragment of actin gene was isolated and idenfied from H.compressa by reverse transcript polymerase chain reaction （RT-PCR）, which contained 773 bp and encoded 257 amino acid. According to the sequence of actin gene above, a quantitative real-time PCR method based on SYBR Green I dye was developed.The quantitative real-time PCR method established in the study had the advantages of high efficiency, wide linear range （R2=0.995,E=101.3%）. The results of this study might provide a basis for actin used as a reference gene to analyze the H.compressa gene quantitatively.