Buchko J, Klassen G R
Department of Microbiology, University of Manitoba, Winnipeg, Canada.
Curr Genet. 1990 Oct;18(3):203-5. doi: 10.1007/BF00318381.
About one-half of the ribosomal repeat unit of two isolates of Pythium ultimum was amplified by means of the polymerase chain reaction using one primer pair. The amplified region includes a small part of the large subunit ribosomal RNA gene, about half of the small subunit ribosomal RNA gene, and the entire intergenic region. The intergenic region of both isolates of P. ultimum has length heterogeneity due to the presence of subrepeat arrays (Klassen and Buchko 1990). PCR amplification of the heterogeneous target DNA resulted in sets of fragments which accurately reflect the heterogeneity in the target DNA, although there is a preferential amplification of the smaller targets. PCR product sizes ranged from 4.6 to 5.8 kb.
利用一对引物通过聚合酶链反应扩增了终极腐霉两个分离株核糖体重复单元的约一半。扩增区域包括大亚基核糖体RNA基因的一小部分、小亚基核糖体RNA基因的约一半以及整个基因间隔区。由于存在亚重复序列阵列,终极腐霉两个分离株的基因间隔区存在长度异质性(克拉森和布赫科,1990年)。对异质靶DNA进行PCR扩增产生了一系列片段,这些片段准确反映了靶DNA中的异质性,尽管较小的靶标存在优先扩增。PCR产物大小范围为4.6至5.8 kb。