Molecular cloning and sequencing of the glycogen phosphorylase gene from Escherichia coli
Molecular cloning and sequencing of the glycogen phosphorylase gene from Escherichia coli
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DOI：
10.1016/0014-5793(89)80128-0
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发表时间：
1989-01
期刊：
影响因子：
3.5
通讯作者：
Yong‐Lark Choi;M. Kawamukai;R. Utsumi;H. Sakai;T. Komano
中科院分区：
文献类型：
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作者：
Yong‐Lark Choi;M. Kawamukai;R. Utsumi;H. Sakai;T. Komano
编码糖原磷酸化酶的glgpgene是从大肠杆菌的基因组文库中克隆出来的。glgpgene的核苷酸序列包含一个单一的开放阅读框，编码一个由790个氨基酸残基组成的蛋白质。glgP基因产物是mr87000的多肽，经SDS -聚丙烯酰胺凝胶电泳证实。推导出的氨基酸序列表明，两者具有同源性。coliand rabbitglgP, humanglgP, potatoglgP, andE。大肠杆菌malp分别为48.6%、48.6%、42.3和46.1%。在这一同源区域内，活性位点、糖原储存位点和吡哆醛- 5′-磷酸结合位点被很好地保守。引入glgpgene多拷贝后，糖原磷酸化酶活性增加。
TheglgPgene, which codes for glycogen phosphorylase, was cloned from a genomic library ofEscherichia coli. The nucleotide sequence of theglgPgene contained a single open reading frame encoding a protein consisting of 790 amino acid residues. The glgP gene product, a polypeptide ofMr87 000, was confirmed by SDS‐polyacrylamide gel electrophoresis. The deduced amino acid sequence showed that homology betweenglgPofE. coliand rabbitglgP, humanglgP, potatoglgP, andE. coli malPwas 48.6, 48.6, 42.3, and 46.1%, respectively. Within this homologous region, the active site, glycogen storage site, and pyridoxal‐5′‐phosphate binding site are well conserved. The enzyme activity of glycogen phosphorylase increased after introduction on a multicopy of theglgPgene.