Harrier L A, Wright F, Hooker J E
Biotechnology Unit, Plant Science Division, Scottish Agricultural College, Kings Buildings, Mains Road, Edinburgh EH9 3JG, Scotland, UK.
Curr Genet. 1998 Dec;34(5):386-92. doi: 10.1007/s002940050411.
The 3-phosphoglycerate kinase (PGK) gene of the arbuscular mycorrhizal fungus Glomus mosseae has been isolated by differential RNA display (DD). Experimentally, the technique of DD was utilized to simultaneously compare the mRNA transcript populations from Lycopersicon esculentum root systems colonized by the arbuscular mycorrhizal fungus G. mosseae and non-mycorrhizal plants. A differentially expressed band was isolated and cloned from mycorrhizal tissue. The isolated DD fragment was screened and sequenced. A reverse transcription-PCR (RT-PCR) technique detected transcripts in germinated spores and G. mosseae-colonized root systems but not in uncolonised root systems. The full-length cDNA was isolated by RACE and corresponded to the 3-phosphoglycerate kinase gene of G. mosseae. The cDNA encodes a polypeptide of 416 amino acids, with a predicted molecular weight of 44 764 Da. The PGK proteins shows high homology to those PGKs of other fungi and the phylogenetic relationship among 14 isolated fungal PGK sequences is illustrated.
通过差异RNA显示（DD）技术分离到了丛枝菌根真菌摩西球囊霉（Glomus mosseae）的3-磷酸甘油酸激酶（PGK）基因。实验中，利用DD技术同时比较了被丛枝菌根真菌摩西球囊霉侵染的番茄根系和非菌根植物的mRNA转录本群体。从菌根组织中分离并克隆出一条差异表达条带。对分离得到的DD片段进行筛选和测序。逆转录聚合酶链反应（RT-PCR）技术检测到该转录本存在于萌发的孢子和被摩西球囊霉侵染的根系中，而在未被侵染的根系中未检测到。通过RACE技术分离得到全长cDNA，其对应于摩西球囊霉的3-磷酸甘油酸激酶基因。该cDNA编码一个由416个氨基酸组成的多肽，预测分子量为44764道尔顿。PGK蛋白与其他真菌的PGK具有高度同源性，并展示了14条分离得到的真菌PGK序列之间的系统发育关系。
