_植物内生源产普鲁兰酶细菌的筛选及其酶活性测定.docVIP

植物内生源产普鲁兰酶细菌的筛选及其酶活性测定 摘 要 作为一种非常重要的药用植物的蒺藜，其具有很多种的药理活性，比如调血脂、降低血压、催欲、抗癌抗菌、利尿等等。现在有很多的研究方向是与蒺藜植株内主要的成分以及药理作用有关。最近几年的研究表明植物的内生菌与植物的许多药用机理有着很紧密的关系。因此本实验就以蒺藜为试验材料，经过用固体分离培养基培养其的根茎组织，分离得到14种细菌，再通过用鉴别培养基筛选出6株产普鲁兰酶的菌株，但只有一株水解圈比较大，对其编号WKX11。为了进一步测定所筛选出来的细菌的产酶情况及其所产酶的活性，本实验用了产酶培养基对试验所筛选出来的细菌所产酶进行了酶活性测定。最后从本实验所培养的细菌菌液中提取DNA，对其DNA进行PCR扩增，然后将试验中获得的PCR产物在电泳液里进行凝胶电泳 SCREENING AND PRODUCTION OF BIOGENIC ACTIVITY ASSAY PULLULANASE BACTERIA IN PLANTS ABSTRACT Tribulus as a very important medicinal plant, which has a wide variety of pharmacological activities, such as adjusting blood fat, lower blood pressure, aphrodisiac, anti-bacterial, diuretic and so on. There are a lot of studies and research on the pharmacological effects of the component within its plant. In recent years many studies have shown that the efficacy of plant endophyte and plants has a very close relationship. Therefore, this experiment Take terrestris as materials, after separation of the solid culture medium with its roots organization, isolated 14 kinds of bacteria, and then screened by differential medium pullulanase producing strain of six, but only one hydrolysis circle is relatively large, their number WKX11. Produced and its activity measured for enzyme production in order to further filter out bacteria, this experiment with the medium on enzyme tests filter out bacteria produced the enzyme activity assay. The last extract from this experiment in broth culture bacterial DNA, PCR amplification of their DNA, and then the PCR product was obtained in the test solution in electrophoresis gel electrophoresis to obtain target band after electrophoresis, the target band after cloning and sequencing were obtained conserved sequence of the target strain; sequencing will get into the NCBI database comparison. The final consolidated morphology, biochemical and molecular biological assay results, the target strain identified as Aerobacter Aerogenes. Having pullulanase producing properties, it can be used as spare prod