桑黄肌动蛋白基因片段的克隆及序列分析

Cloning and sequence analysis of Actin gene fragment from Phellinus baummi

目的对桑黄肌动蛋白(Actin)基因进行克隆及序列分析。方法搜索网上已知数据库中Actin基因,与本实验室测得的桑黄转录组数据进行生物信息学比对,找到转录组数据中与Actin相似性最高的片段,并根据其设计引物,提取桑黄菌丝体总RNA为模板,采用RT-PCR的方法扩增Actin基因片段并对PCR产物进行测序。结果得到一段839 bp的序列,序列分析表明,该片段编码279个氨基酸,与其他物种Actin基因核苷酸序列同源性在87%以上。结论首次从桑黄中克隆出了Actin基因,为有效利用该基因奠定了基础。

Objective To clone and analyze the cDNA fragment encoding Actin gene ofPhellinus baummi. Methods A pair of primers were designed based on the transcriptome data ofP baummi after comparing with Actin gene of other species. Taking total RNA from P. baummi as template, Actin gene fragment was obtained by reverse transcription polymerase chain reaction （RT-PCR） and the PCR products were sequenced. Results The Actin gene fragment from P. baummi contained 839 bp, encoding 279 amino acids. The sequence analysis suggested that the nucleotide sequence shared over 87% of homology with Actin gene from other species. Conclusion It is first reported that a novel Act-in gene is cloned from P. baummi. This work lays a base for the effective utilization ofActin gene.