FIREFLY LUCIFERASE AS A REPORTER ENZYME FOR MEASURING GENE-EXPRESSION IN VEGETATIVE AND SYMBIOTIC RHIZOBIUM-MELILOTI AND OTHER GRAM-NEGATIVE BACTERIA
FIREFLY LUCIFERASE AS A REPORTER ENZYME FOR MEASURING GENE-EXPRESSION IN VEGETATIVE AND SYMBIOTIC RHIZOBIUM-MELILOTI AND OTHER GRAM-NEGATIVE BACTERIA
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DOI：
10.1016/0378-1119(89)90336-3
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发表时间：
1989-09-01
期刊：
影响因子：
3.5
通讯作者：
HELINSKI, DR
中科院分区：
文献类型：
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作者：
PALOMARES, AJ;DELUCA, MA;HELINSKI, DR
携带北美萤火虫荧光素酶基因（luc） cDNA拷贝的DNA片段融合到.lambda。pR启动子在大肠杆菌中的表达[de Wet et al., Proc. Natl.]学会科学。USA 82(1985) 7870-7873]，插入到广泛宿主质粒载体中，并在多种革兰氏阴性菌中建立。荧光素酶活性，由。lambda。pR启动子，在完整细胞和从melilotirhizobium， R. phaseoli， R. fredii, Pseudomonas aeruginosa, Agrobacterium tumefaciens， accoaceticus不动杆菌和Azotobacter vinelandii提取的细胞中均检测到。通过对完整细胞和提取物的测量，观察到铜绿假单胞菌和三种根瘤菌的最高活性水平。荧光素酶活性在成熟紫花苜蓿根瘤中的表达量也相对较高。用x射线胶片或纯化的类细菌提取物很容易在完整的结节中检测到这种活性。
A DNA segment carrying a cDNA copy of the luciferase gene (luc) of the North American firefly Photinus pyralis, fused to the .lambda. pR promoter and expressed in Escherichia coli [de Wet et al., Proc. Natl. Acad. Sci. USA 82 (1985) 7870-7873], was inserted into a broad-host-range plasmid vector and established in a variety of Gram-negative bacteria. Luciferase activity, expressed from the .lambda. pR promoter, was detected in both intact cells and extracts prepared from cells of strains of Rhizobium meliloti, R. phaseoli, R. fredii, Pseudomonas aeruginosa, Agrobacterium tumefaciens, Acinetobacter calcoaceticus and Azotobacter vinelandii. The highest levels of activity, determined by measurements of both intact cells and extracts, were observed for P. aeruginosa and the three species of Rhizobium examined. Expression of luciferase activity also was relatively high in R. meliloti bacteroids of mature alfalfa nodules. This activity was readily detectable in intact nodules using x-ray film or in extracts prepared from purified bacteroid.