Cloning, sequence analysis and expression in Escherichia coli of the gene encoding a uricase from the yeast-like symbiont of the brown planthopper, Nilaparvata lugens
Cloning, sequence analysis and expression in Escherichia coli of the gene encoding a uricase from the yeast-like symbiont of the brown planthopper, Nilaparvata lugens
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DOI：
10.1016/s0965-1748(99)00116-2
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发表时间：
2000-02-01
影响因子：
3.8
通讯作者：
Ishikawa, H
中科院分区：
文献类型：
--
作者：
Hongoh, Y;Sasaki, T;Ishikawa, H
克隆了褐飞虱（Nilaparvata lugens）酵母样真菌内共生体的尿酸氧化酶（uricase; EC 1.7.3.3）基因及其侧翼序列。该基因包含一个987 bp的开放阅读框架，该框架被一个96 bp的内含子分成两部分。该基因编码296个氨基酸，与黄曲霉尿酸酶的同源性为62%。推导的分子量为32，882，预测的等电点为6.06。该共生体的尿酸酶保守所有已知的一致基序，除了C-末端PTS-1，Ser-basic-Leu。PTS-1第三位的亮氨酸被共生体尿酸酶C-末端的丝氨酸取代。该共生菌的尿酸酶基因在大肠杆菌中成功表达，并纯化了带有组氨酸残基标签的产物。共生体的尿酸酶，从而产生的，是活跃的植物和动物，但不如那些从其他真菌。(C)2000爱思唯尔科技有限公司版权所有。
A urate oxidase (uricase; EC 1.7.3.3) gene of the yeast-like fungal endosymbiont of the brown planthopper, Nilaparvata lugens, was cloned, and sequenced together with its flanking regions. The gene comprised a open reading frame of 987 bp, that was split into two parts by a single 96 bp intron. The encoded uricase was 296 amino acids with 62% sequence identity with that of Aspergillus flavus. The molecular weight deduced was 32,882, and the predicted isoelectric point was 6.06. The symbiont's uricase conserved all the known consensus motifs, except the C-terminal PTS-1, Ser-basic-Leu. The leucine at the third position of PTS-1 was replaced by serine in the C-terminus of the symbiont's uricase. The symbiont's uricase gene was successfully expressed in Escherichia coli, and the product, tagged with histidine residues, was purified. The symbiont's uricase, thus produced, was as active as those from plants and animals, but less active than those from other fungi. (C) 2000 Elsevier Science Ltd. All rights reserved.