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机构地区:[1]中国热带农业科学院热带生物技术研究所,海南海口571101 [2]海南医学院,海南海口571101
出 处:《热带作物学报》2011年第8期1512-1517,共6页Chinese Journal of Tropical Crops
基 金:国家自然科学基金(30960032);中央级公益性科研院所基本科研业务费专项资金资助项目(ITBBZX0841;ITTBZD09-41)
摘 要:以微茫藻为材料,利用抑制差减杂交技术,分离鉴定了10种与已知功能基因具有较高同源性的基因片段,这些片段的同源基因均与氮源的吸收利用相关。实时荧光定量PCR验证了7种基因在氮缺乏条件下转录水平明显上调,包括:硝酸还原酶基因、NMT1/THI5家族蛋白基因、N-羟甲基-L-丙氨酸脱氢酶基因、核糖体蛋白L36a基因、FAD/NAD(P)结合的氧化还原酶基因、NADH:细胞色素b5还原酶基因和细胞色素cd1-亚硝酸还原酶类似基因。而天冬氨酸转氨酶、红细胞膜蛋白7.2b和γ-谷氨酰磷酸还原酶这3种基因在氮缺乏时下调表达。To elucidate the effects of nitrogen starvation on the expression of nitrogen assimilation related genes, ten ESTs were isolated in Micractinium pusillum by using Suppression Subtraction Hybridization (SSH) , which showed high similarity to those genes involved in nitrogen assimilation. Real-time PCR confirmed that the transcript levels of seven genes were significantly increased after nitrogen starvation, including genes encoding nitrate reductase, NMT1/HI5 like domain protein, strombine dehydrogenase, oxidoreductase FAD/NAD(P)-binding, ribosomal proein L36a, cytochrome cdl-nitrite reductase-like and NADH:cytochrome b5 reductase. On the other hand, genes encoding aspartate aminotransferase, stomatin and gamma-glutamyl phosphate reductase were down- regualted by nitrogen starvation in Micractinium pusillum.
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