Brown A E, Muthumeenakshi S, Sreenivasaprasad S, Mills P R, Swinburne T R
Department of Applied Plant Science, Queen's University of Belfast, UK.
FEMS Microbiol Lett. 1993 Mar 15;108(1):117-20. doi: 10.1111/j.1574-6968.1993.tb06083.x.
An oligonucleotide primer (ChInt) was synthesised from the variable internally transcribed spacer (ITS) 1 region of ribosomal DNA (rDNA) of Cylindrocarpon heteronema. PCR with primers ChInt and ITS4 (from a conserved sequence of the rDNA) amplified a 470-bp fragment from several isolates of C. heteronema but not from various apple wood saprophytes. Amplification of this fragment was achieved from 1-2 pg of fungal DNA. These primers amplified a fragment of the same size from DNA extracted from cankered wood but only after impurities were removed from the DNA on a Qiagen tip-5 column. Southern hybridization analysis confirmed the 470-bp fragment from C. heteronema DNA and cankered wood to be identical.
从柱孢属异形根霉菌核糖体DNA（rDNA）的可变内部转录间隔区（ITS）1区域合成了一个寡核苷酸引物（ChInt）。用引物ChInt和ITS4（来自rDNA的保守序列）进行PCR，从多个异形根霉菌分离株中扩增出一个470bp的片段，但从各种苹果木腐生菌中未扩增出该片段。从1-2pg的真菌DNA中实现了该片段的扩增。这些引物从溃疡木提取的DNA中扩增出相同大小的片段，但前提是在Qiagen tip-5柱上从DNA中去除杂质之后。Southern杂交分析证实，来自异形根霉菌DNA和溃疡木的470bp片段是相同的。
