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浑球红细菌谷氨酸合酶基因(glt)的克隆和图谱分析 被引量:3
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作者 鲁涛 吴永强 宋鸿遇 《微生物学报》 CAS CSCD 北大核心 1996年第1期1-11,共11页
利用转座子Tn5随机插入诱变筛选得到12株浑球红细菌(Rhodobacter sphaeroides)氨同化缺陷突变株(Asm^-)。这些突变株胞内均无GOGAT活性,同时它们均无固氮酶活性(Nif^-),并且具有氮代谢多效性缺失表型(Ntr^-)。将含有Azorhizobium sesban... 利用转座子Tn5随机插入诱变筛选得到12株浑球红细菌(Rhodobacter sphaeroides)氨同化缺陷突变株(Asm^-)。这些突变株胞内均无GOGAT活性,同时它们均无固氮酶活性(Nif^-),并且具有氮代谢多效性缺失表型(Ntr^-)。将含有Azorhizobium sesbaniae ORS571的完整glt基因的质粒pHB10转入突变株中能互补上述表型。通过筛选携带Tn5的R-prime质粒克隆了glt::Tn5片段。Southern杂交证明所克隆glt::Tn5片段与E. coli的gltBD基因有同源性。用此片段与以pLAFR3为载体所构建的R. sphaeroides 601基因文库进行菌落原位杂交筛选到了携带glt基因的cosmid pLT27。pLT27能互补所有12株R.sphaeroides氨同化缺陷突变株。酶切分析表明在该cosmid中插人的染色体DNA片段大小约为26.5kb。以pRK415为载体亚克隆了4.0kb与10.5kh的pLT27的Hindlll酶切片段,分别命名为pLTRK271与pLTRK272。pLTRK272能互补变种GT6、GT10、GT11,pLTRK271能互补其余的变种。通过单、双酶切作出了pLTRK271与pLTRK272的物理图谱。上述10.5kb的pLT27Hindlll酶切片段与E. coil gltBD基因具有同源性。Southern杂交分析表明Tn5均插在变种染色体的同一区域并得出了其插入位点。由此可确定glt基因大致位于一段11kb的EcoRl-Hindlll克隆片段内。 展开更多
关键词 浑球红细菌 谷氨酸合酶基因 基因克隆 图谱分析
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浑球红细菌谷氨酸合酶大亚单位基因(gltB)的序列分析 被引量:1
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作者 鲁涛 吴永强 宋鸿遇 《微生物学报》 CAS CSCD 北大核心 1997年第2期87-94,共8页
测定了浑球红细菌(Rhodobacter sphaeroides)谷氨酸合酶大亚单位基因(gltB)及其5'端和3'端的序列,全长为5510bp。序列分析表明,R. sphaeroides gltB基因全长为4636bp。从核苷酸序列推测其蛋白质分子量约为164kD。R. sphaeroides... 测定了浑球红细菌(Rhodobacter sphaeroides)谷氨酸合酶大亚单位基因(gltB)及其5'端和3'端的序列,全长为5510bp。序列分析表明,R. sphaeroides gltB基因全长为4636bp。从核苷酸序列推测其蛋白质分子量约为164kD。R. sphaeroides gltB基因与Azospirillum brasilense和Escherichia coli的gltB基因DNA序列有很高的同源性。其蛋白质氨基酸序列与A. brasilense gltB基因产物GltB也具有很高的同源性。此外,还对R. sphaeroides GltB的各可能功能区进行了分析,发现它们具有很高的保守性。 展开更多
关键词 浑球红细菌 谷氨酸合酶基因 DNA序列
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Suppression of glutamate synthase genes significantly affects carbon and nitrogen metabolism in rice (Oryza sativa L.) 被引量:13
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作者 LU YongEn LUO Feng +2 位作者 YANG Meng LI XiangHua LIAN XingMing 《Science China(Life Sciences)》 SCIE CAS 2011年第7期651-663,共13页
Rice (Oryza sativa) glutamate synthase (GOGAT,EC 1.4.1.14) enzymes have been proposed to have great potential for improving nitrogen use efficiency,but their functions in vivo and their effects on carbon and nitrogen ... Rice (Oryza sativa) glutamate synthase (GOGAT,EC 1.4.1.14) enzymes have been proposed to have great potential for improving nitrogen use efficiency,but their functions in vivo and their effects on carbon and nitrogen metabolism have not been systematically explored.In this research,we analyzed transcriptional profiles of rice GOGAT genes using a genome-wide microarray database,and investigated the effects of suppression of glutamate synthase genes on carbon and nitrogen metabolism using GOGAT co-suppressed rice plants.Transcriptional profiles showed that rice GOGAT genes were expressed differently in various tissues and organs,which suggested that they have different roles in vivo.Compared with the wild-type,tiller number,total shoot dry weight,and yield of GOGAT co-suppressed plants were significantly decreased.Physiological and biochemical studies showed that the contents of nitrate,several kinds of free amino acids,chlorophyll,sugars,sugar phosphates,and pyridine nucleotides were significantly decreased in leaves of GOGAT co-suppressed plants,but the contents of free ammonium,2-oxoglutarate,and isocitrate in leaves were increased.We conclude that GOGATs play essential roles in carbon and nitrogen metabolism,and that they are indispensable for efficient nitrogen assimilation in rice. 展开更多
关键词 glutamate synthase Oryza sativa CO-SUPPRESSION carbon and nitrogen metabolism
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