天麻ITS序列的聚合酶链反应扩增

The Amplification of ITS Sequence of Gastrodia elata with Polymerase Chain Reaction

目的:为天麻核糖体DNA中的内转录间隔区(internal transcribed spacer,ITS)序列分析提供可靠、特异的聚合酶链反应(PCR)条件。方法:选择ITS序列通用引物对天麻ITS序列进行PCR扩增,对一系列PCR条件参数进行摸索,同时对PCR产物进行纯化后直接测序。结果:天麻ITS序列PCR扩增产物约为750 bp,不同来源样本存在有一定的差异。经测序后与Genebank中相关序列比对,天麻ITS序列总长度为598 bp,其中ITS1为235 bp,ITS2为209 bp,5.8S为154 bp,G+C(%)为52%。结论:扩增天麻ITS序列的PCR反应条件可以得到可靠、特异的结果。

Objective： To establish a reliable and specific PCR method for the amplification of the internal transcribed spacer （ITS） region of nuclear ribosomal DNA of Gastrodia elata. Methods： The ITS sequence was amplified by PCR with universal primers of ITS. The PCR parameters for ITS sequence amplification were optimized and PCR product was directly sequenced after purification. Results： The PCR product was about 750 bp. There occurred little diversity among the samples from different sources. After sequenced and compared with the optimized sequences in Genbank, the total length of ITS sequence of Gastrodia elata was determined as 598 bp including 235 bp of ITS1,209 bp of ITS2, and 153 bp of 5.8 S, and the （G ＋C）% was 52%. Conclusions： With the optimized PCR conditions, the amplification of ITS sequence of Gastrodia elata can produce reliable and specific results.