应用发光酶基因对快生型大豆根瘤菌HN01结瘤作用进行检测  

luxAB GENES AS MARKER FOR DETECTING RHIZOBIUM FREDII HN01 NODULATION FUNCTIONS

作　　者：莫才清[1] 覃雅丽[1] 周俊初[1] 李阜棣[1] 

机构地区：[1]华中农业大学微生物系,武汉430070

出　　处：《微生物学报》1998年第3期213-218,共6页Acta Microbiologica Sinica

基　　金：国家高技术863资助;国家自然科学基金

摘　　要：含发光酶基因luxAB的Tn5转座子自杀质粒pHNC3，在辅助质粒pRK2013的帮助下，转入快生型大豆根瘤菌HN01中小，pHNC3经自杀重组，其Tn5－luxAB转座插入HN01基因组中，从而赋予HN01以发光活性。挑取具有发光活性的HN01杂交单菌落，进行质粒快检和以luxAB为探针的分子杂交，选取Tn5-luxAB分别插入到HN01染色体上和不同质粒上的标记菌株，进行灭菌盆栽实验，并对一株Tn5-luxAB标记于染色体上的菌株HN01LC02进行了模拟大豆栽培条件下的有菌盆栽实验，包括对发光根瘤菌占瘤率的测定和发光根瘤在根系上分布情况的测定。A suicide plasmid PHNC3 which contains Tn5-luxAB was transfered into Rhizobium fredii HN01 by the help of pRK2013. Then Tn5-luxAB inserted on the genenome of HN01 and gave luminescence activity. The luminescence colonies were picked up and the Eckhardt gel was performed for plasmids profile detection. The location of Tn5-luxAB on the genenome was determined using the luxAB as probe.The colonies which were marked by Tn5-luxAB on the chromosome and different plasmids of HN01 were choosed for pot experiment and a chromosome labelled strain HN01LC02 was detected by soil pot experiment The detections included the nodulation occupancy and the luminescent nodules distribution on the root system formed by the luxAB-marked rhizobia.

关 键 词：发光酶基因 大豆根瘤菌 检测 结瘤 

分 类 号：Q939.114[生物学—微生物学] Q945.13