Cloning, characterization, and nucleotide sequence analysis of a Zymomonas mobilis phosphoglucose isomerase gene that is subject to carbon source-dependent regulation
Cloning, characterization, and nucleotide sequence analysis of a Zymomonas mobilis phosphoglucose isomerase gene that is subject to carbon source-dependent regulation
复制标题
受碳源依赖性调控的运动发酵单胞菌磷酸葡萄糖异构酶基因的克隆、表征和核苷酸序列分析
作者：
T. Hesman;W. O. Barnell;T. Conway
通过对大肠杆菌磷酸葡萄糖异构酶（pgi）突变体的基因互补，克隆了运动发酵单胞菌的pgi基因。酶分析和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳证实了在E.大肠杆菌中携带Z. mobilis pgi基因。pgi基因在Z上仅以一个拷贝存在。mobilis基因组pgi区的核苷酸序列分析揭示了一个1,524 bp的开放阅读框架，前面是一个强的Shine-Dalgarno序列。pgi基因编码507个氨基酸的蛋白质，预测分子量为55，398。Z. mobilis磷酸葡萄糖异构酶与来自其它物种的酶具有38 - 43%的同一性。北方（RNA）杂交分析表明，pgi转录产物长度为1.8kb。发现pgi转录水平受生长阶段以及碳源和能源的影响。转录水平增加，相对于总RNA在对数生长，果糖比葡萄糖生长时高三倍。这些转录水平的变化改变了培养物中磷酸葡萄糖异构酶的活性。差异mRNA的稳定性不是一个因素，因为在葡萄糖生长的细胞中，pgi转录物的半衰期为6.3分钟，在果糖生长的细胞中为6.0分钟。因此，转录速率的增加似乎是至少部分负责增加磷酸葡萄糖异构酶水平观察Z。在果糖上生长的mobilis。
The Zymomonas mobilis gene encoding phosphoglucose isomerase (pgi) was cloned by genetic complementation of an Escherichia coli pgi mutant. An enzyme assay and sodium dodecyl sulfate-polyacrylamide gel electrophoresis confirmed the presence of excess amounts of phosphoglucose isomerase in E. coli clones carrying the Z. mobilis pgi gene. The pgi gene is present in only one copy on the Z. mobilis genome. Nucleotide sequence analysis of the pgi region revealed an open reading frame of 1,524 bp preceded by a strong Shine-Dalgarno sequence. The pgi gene encodes a 507-amino-acid protein with a predicted molecular weight of 55,398. Z. mobilis phosphoglucose isomerase is between 38 and 43% identical to the enzyme from other species. Northern (RNA) blot analysis showed that the pgi transcript is 1.8 kb in length. The level of the pgi transcript was found to be influenced by the phase of growth and by the carbon and energy sources. Transcript levels increased with respect to total RNA during logarithmic growth and were threefold higher when grown on fructose than on glucose. These changes in transcript levels paralleled phosphoglucose isomerase activities in the cultures. Differential mRNA stability was not a factor, since the half-life of the pgi transcript was 6.3 min in glucose-grown cells and 6.0 min in fructose-grown cells. Thus, an increase in the rate of transcription appears to be at least partially responsible for the increased levels of phosphoglucose isomerase observed for Z. mobilis grown on fructose.
影响因子：
56.9
作者：
GURNEY, ME;HEINRICH, SP;YIN, HS
通讯作者：
YIN, HS
葡萄糖磷酸异构酶活性位点组氨酸的亲和标记和表征。
DOI：
--
发表时间：
1980
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Gibson,DR;Gracy,RW;Hartman,FC
通讯作者：
Hartman,FC