来自禾本科布氏白粉菌（一种“顽固”真菌）的羊毛甾醇14α-脱甲基酶（CYP51）基因的PCR克隆及点突变检测
PCR cloning and detection of point mutations in the eburicol 14alpha-demethylase (CYP51) gene from Erysiphe graminis f. sp. hordei, a "recalcitrant" fungus.
作者信息
Délye C, Bousset L, Corio-Costet M F
机构信息
Unité de Recherches Intégrées sur la Vigne, Institut National de la Recherche Agronomique, Domaine de la Grande Ferrade, B. P. 81, F-33883 Villenave d'Ornon cédex, France.
出版信息
Curr Genet. 1998 Dec;34(5):399-403. doi: 10.1007/s002940050413.
Molecular studies of some micro-organisms are hampered by the difficulty of obtaining sufficient amounts of nucleic acids. A cloning strategy based on PCR has therefore been used to clone the eburicol 14alpha-demethylase (CYP51) gene of the obligate fungus Erysiphe graminis f. sp. hordei (Egh) using minute amounts of genomic DNA. The CYP51 gene encodes the enzymatic target of a major group of fungicides. Sequencing CYP51 from different Egh isolates revealed the occurrence of two alleles for this gene. An allele-specific PCR assay was developed to detect each CYP51 allele.
一些微生物的分子研究因难以获得足够量的核酸而受到阻碍。因此，一种基于聚合酶链式反应（PCR）的克隆策略已被用于克隆专性真菌大麦白粉病菌（Erysiphe graminis f. sp. hordei，Egh）的羊毛甾醇14α-脱甲基酶（CYP51）基因，该克隆过程仅使用微量的基因组DNA。CYP51基因编码了一类主要杀菌剂的酶作用靶标。对不同Egh分离株的CYP51进行测序，结果显示该基因存在两个等位基因。随后开发了一种等位基因特异性PCR检测方法来检测每个CYP51等位基因。
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