【摘要】 [目的]对甜杨葡萄糖-6磷-酸脱氢酶(G6PDH)进行克隆及功能预测。[方法]采用电子克隆和RT-PCR相结合的方法首次从甜杨中分离了G6PDH基因,通过B last和其他生物信息学软件进行功能预测。[结果]甜杨G6PDH基因的cDNA长1 697 bp,可编码510个氨基酸,cDNA序列及其推导的氨基酸序列均与其他植物G6PDH存在着较高的同源性,说明所获得的cDNA是甜杨G6PDH基因(PsG6PDH,AY445917)。同时,运用PCR扩增技术获得了甜杨G6PDH基因组序列,测序结果表明,基因组DNA长度为5 040 bp,含有15个外显子和14个内含子。Southern杂交结果表明,G6PDH基因在甜杨的基因组中可能是单拷贝或者低拷贝。通过网络服务器平台进行PsG6DH的功能预测,结果显示,PsG6PDH为G6PDH的成员之一,含有NADP结合区和G6P结合区2个保守功能域,具有多个磷酸化位点和跨膜区域,但不含有信号肽,表明PsG6DH可能作为膜受体而起作用。另外,G6PDH的电子表达谱分析结果发现,G6PDH在多种组织和不同发育过程均表达,并涉及各种逆境胁迫应答反应,这也说明了G6PDH在植物生长反育中的重要地位。[结论]可为下一步研究PsG6PDH基因的分子功能提供参考。更多还原

【Abstract】 [Objection]In silico cloning and function prediction of glucose-6-phosphate dehydrogenase gene(G6PDH) was researched in the paper.[Method] The G6PDH gene was cloned from P.suaveolens by electronic PCR and RT-PCR,and function prediction was carried out by Blast and other bioinformatics softs.[Result] The full length PsG6PDH was 1 697 bp and encoded 510 amino acids.cDNA and amino acid sequences showed high identity with other plant G6PDH genes.P.suaveolens(fragrant poplar) G6PDH genomic DNA was isolated by PCR and its sequence contained 15 exons and 14 introns.In addition,Southern analysis revealed that PsG6PDH is single or low copies in the fragrant poplar genome.Function prediction showed that PsG6PDH was a new member of plant G6PDH family,containing NADP-binding domain and G6P-binding domain,with some phosphorylation sites.It had no signal peptide,but one trasmembrane domain in N terminal,indicating that PsG6PDH was acted as a receptor in membrane.Moreover,electronic expression profiles revealed that G6PDH expressed in whole plants and varied development stage,and involved in different abiotic stresses,suggesting that G6PDH played an important role in the growth and development of plants.[Conclusion] The report provided important information for the further molecular function research of PsG6PDH.更多还原