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The insect\|resistant activity of Bt cotton has significant variation in different tissue and growing period. Bioassay, the method used to determine the insect\|resistant activity of Bt cotton to bollworm inside the country, can not determine the expression levels of Bt protein. The ELISA method established by us resolves this problem. Compared with bioassay, ELISA has the merits of spending shorter time, quantifying Bt toxin protein and easy operation. We find that Bt cotton has different Bt toxin...

The insect\|resistant activity of Bt cotton has significant variation in different tissue and growing period. Bioassay, the method used to determine the insect\|resistant activity of Bt cotton to bollworm inside the country, can not determine the expression levels of Bt protein. The ELISA method established by us resolves this problem. Compared with bioassay, ELISA has the merits of spending shorter time, quantifying Bt toxin protein and easy operation. We find that Bt cotton has different Bt toxin protein content in different tissue and growing period using ELISA method. Leaves and flower petals contain more Bt toxin protein than boll and bracteal leaves. In the late growing period, Bt toxin protein content of every tissue decreases significantly.

转苏云金杆菌 (Bacillusthuringensis)杀虫蛋白基因棉花 (下称Bt棉 )在不同生长期及不同器官上对棉铃虫杀虫活性存在明显差异。目前国内普遍使用生物测试法检测Bt棉对棉铃虫的杀虫活性 ,但无法定量检测Bt棉中杀虫蛋白的表达量。本实验室建立的酶联免疫学测定方法(ELISA) ,可定量检测棉器官内杀虫蛋白的含量。它具有检测周期短 ,操作简便的优点。用ELISA方法检测Bt棉杀虫蛋白的含量 ,结果表明 ,棉叶和花瓣杀虫蛋白含量最高 ,铃和蕾次之。在棉株生长的后期 ,各器官杀虫蛋白的含量则有大幅度的下降。

By applying antibody sandwich enzyme linked immunosorbent assay method,the content of Bt (Bacillus thuringiensis Berliner) toxin protein in the Bt transgenic cotton was tested. The results showed that Bt toxin protein was detected in all the test organs of the transgenic cotton ,but its contents varied significantly among different organs in different stages. The highest content occurred in the fully expanded leaves in seedling stage, while those in root , stem , petiole in the same stage were...

By applying antibody sandwich enzyme linked immunosorbent assay method,the content of Bt (Bacillus thuringiensis Berliner) toxin protein in the Bt transgenic cotton was tested. The results showed that Bt toxin protein was detected in all the test organs of the transgenic cotton ,but its contents varied significantly among different organs in different stages. The highest content occurred in the fully expanded leaves in seedling stage, while those in root , stem , petiole in the same stage were rather low. In flowering boll stage , much higher content occurred in ovary on the day of flowering ,but those in pistil and stamen were lower , and those in petal and bract were the lowest .There were significant differences of Bt toxin protein contents among the functional leaves in different stages, the order of content was seedling>quaring>flowering boll. Consequently, The expression of Bt toxin protein in Bt transgenic cotton decreased with advance of development .It shoud be the reason why the resistance of Bt transgenic cotton to cotton bollworm (Helicoverpa armigera (Hübner)) changed temporally and spatially in cotton developmental processes.

采用抗体夹心 EL ISA技术 ,对转 Bt基因抗虫棉植株中 Bt毒蛋白含量进行了测定。结果表明 ,Bt基因在所有检测到的器官中均有表达 ,但是不同器官中的 Bt毒蛋白含量明显不同。在苗期全展功能叶中 Bt毒蛋白含量最高 ,根、茎和叶柄中 Bt毒蛋白含量较低 ;在花铃期当日开花的子房中 Bt毒蛋白含量较高 ,雌雄蕊中 Bt毒蛋白较低 ,花瓣及苞叶 Bt毒蛋白含量最低。表明 Bt基因在不同器官中的表达强度存在差异。不同生育期的功能叶中 Bt毒蛋白含量差异显著 ,Bt毒蛋白含量在苗期叶片中最高 ,蕾期次之 ,花铃期最低。随着棉花生长发育进程的推进 ,Bt基因在叶片中的表达强度逐渐减弱。Bt基因在棉花体内的表达随着器官的不同、生育时期的不同而表现出时空动态变化。这可能是人们所观察到的转 Bt基因抗虫棉对棉铃虫抗性呈时空动态变化的根本原因。

Improved histochemical staining for GUS activity was used to detect the segregation of transgenes in the different generations of Bt rice crossed to conventional rice varieties. Mendelian segregation was observed in F_2, BC_1 and BC_1F_2 progenies, Thus demonstrating that transgenes inherit as a single dominant gene in the progenies of Bt rice crossed to conventional rice varieties. Western blotting analyses indicate that population plants in the different generations of Bt rice crossed to conventional rice...

Improved histochemical staining for GUS activity was used to detect the segregation of transgenes in the different generations of Bt rice crossed to conventional rice varieties. Mendelian segregation was observed in F_2, BC_1 and BC_1F_2 progenies, Thus demonstrating that transgenes inherit as a single dominant gene in the progenies of Bt rice crossed to conventional rice varieties. Western blotting analyses indicate that population plants in the different generations of Bt rice crossed to conventional rice varieties express higher Bt toxin proteins than that of original Bt rice. Agronomic trait analysis found that there are no significant difference between resistant-plant and susceptible-plant in the progenies of either indica/japonica crosses or japonica/japonica crosses, thus sho wing exciting prospect for Bt rice breeding.

利用GUS反应检测了Bt转基因粳稻(克螟稻)与灿稻、粳稻杂交的不同世代群体的叶片染色情况,结果发现粳粳交F_2出现抗性株与非抗性株的3∶1的分离,籼粳交的BC_1出现1∶1分离,籼粳交的BCF_2出现3∶1分离,表明转基因在杂交后代的传递属单基因显性遗传。利用Western点杂交技术结果发现F_1、F_2和BC_1代的Bt蛋白表达量均超过转基因株供体,呈现出表达能力上的杂种优势。农艺性状考察结果发现,无论是籼粳交,还是粳粳交后代,两者在株高、穗长、单株分蘖数、播始历期和千粒重上无显著差异。这无疑为Bt水稻的育种利用提供了良好的证据。

 
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