以黄瓜品种露丰为材料,根据已报道的6-磷酸葡萄糖酸脱氢酶基因(6PGDH)的保守氨基酸序列设计简并引物,利用RT-PCR技术获得黄瓜6PGDH的cDNA同源片段,命名为CSPG(登录号为EU815934)。该片段长度为1 207 bp,包含一个936 bp的开放阅读框(编码311个氨基酸)和271 bp的Poly A 3′非翻译区末端,无内含子;该基因编码的氨基酸序列与拟南芥、大豆、水稻、玉米、菠菜的6PGDH基因有75%以上的同源性。运用半定量RT-PCR技术对6PGDH基因的转录水平进行分析,结果表明:该基因在叶、根、茎中均有表达,高温胁迫下的表达量高于常温对照,说明6PGDH基因与热胁迫相关。
With homology-based PCR strategy,a fragment cDNA named CSPG(accession number EU815934) was cloned from Cucumis sativus‘Lufeng'.The length of CSPG was 1 207 bp,which encompassed an open reading frame(ORF) with 936 bp encoding 311 amino acid residues and 271 bp non-translation 3'-terminal end region,and no intron existed in CSPG.The protein encoded by CSPG was more than 75%highly homologous 6-phosphogluconate dehydrogenase gene(6PGDH) of Arabidopsis thaliana, Glycine max,Oryza sativa,Zea mays,Spinacia oleracea.The expression of 6PGDH was analyzed by semi-quantitative RTPCR indicating that the gene had expression in leaf,root and stem,and that the amount of expression was higher in heat stress than normal temperature.Which showed that expression of 6PGDH was related to heat stress.