DEVELOPMENT OF PRIMER SETS DESIGNED FOR USE WITH THE PCR TO AMPLIFY CONSERVED GENES FROM FILAMENTOUS ASCOMYCETES
DEVELOPMENT OF PRIMER SETS DESIGNED FOR USE WITH THE PCR TO AMPLIFY CONSERVED GENES FROM FILAMENTOUS ASCOMYCETES
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DOI：
10.1128/aem.61.4.1323-1330.1995
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发表时间：
1995-04-01
影响因子：
4.4
通讯作者：
DONALDSON, GC
中科院分区：
文献类型：
--
作者：
GLASS, NL;DONALDSON, GC
我们构建了9套寡核苷酸引物的基础上，从粗糙脉孢菌和构巢曲霉的基因组的选择丝状子囊菌和半知菌（与丝状子囊菌隶属关系）的基因组的DNA杂交的结果。设计了九组引物来扩增跨越保守基因中的一个或多个内含子的DNA片段。用来自子囊菌、半知菌、担子菌和植物的基因组DNA的9个引物组进行PCR DNA扩增，结果显示其中5个引物组仅从丝状子囊菌和半知菌的DNA扩增出产物。这5对引物是从N.组蛋白3、组蛋白4、β-微管蛋白和质膜ATP酶的crassa基因。利用这5对引物，在丝状子囊菌扩增产物的大小和限制性酶切位点上均观察到多态性。这里描述的引物组可以提供有用的工具，在丝状子囊菌和半知菌（子囊菌隶属）的系统发育研究和基因组分析，以及通过PCR快速区分真菌物种。
We constructed nine sets of oligonucleotide primers on the basis of the results of DNA hybridization of cloned genes from Neurospora crassa and Aspergillus nidulans to the genomes of select filamentous ascomycetes and deuteromycetes (with filamentous ascomycete affiliations). Nine sets of primers were designed to amplify segments of DNA that span one or more introns in conserved genes. PCR DNA amplification with the nine primer sets with genomic DNA from ascomycetes, deuteromycetes, basidiomycetes, and plants revealed that five of the primer sets amplified a product only from DNA of the filamentous ascomycetes and deuteromycetes. The five primer sets were constructed from the N. crassa genes for histone 3, histone 4, beta-tubulin, and the plasma membrane ATPase. With these five primer sets, polymorphisms were observed in both the size of and restriction enzyme sites in the amplified products from the filamentous ascomycetes. The primer sets described here may provide useful tools for phylogenetic studies and genome analyses in filamentous ascomycetes and deuteromycetes (with ascomycete affiliations), as well as for the rapid differentiation of fungal species by PCR.