摘要
通过农杆菌介导法将杀虫晶体蛋白基因cry1Ab导入高粱恢复系115中,共获得13个独立的转基因株系,26株转基因植株,转化率为5.1%;GUS活性、PCR、Southern和Western杂交分析表明,此基因已整合进高粱基因组并得到正确表达。利用ELISA试剂盒测定Cry1Ab蛋白含量,结果表明,不同转基因植株的Cry1Ab蛋白含量有明显差异,最高可达0.850μg/g,占可溶性蛋白的0.016%,还有一些转基因植株中不能表达;同一转基因植株的不同组织中表达量有明显差异,其顺序为:叶>颖壳>籽粒>根>茎;不同部位的叶片也有差异,其顺序为:中部叶>基部叶>新叶。
The insecticidal Bacillus thuringiensis （Bt） cry 1 Ab gene was introduced into sorghum restoring line 115 efficiently mediated by Agrobacterium tumefaciens. A total of 13 independent transgenic plant lines and 26 transgenic plants were regenerated with the average transformation efficiency of 5.1%. The results of GUS expression, PCR analysis, Southern and Western hybridization showed that the crylAb gene had been integrated and expressed exactly in sorghum. The expression level of cry lAb gene was detected in transgenic sorghum plants by using ELISA kits. There were obvious differences in CrylAb protein content with the different transgenic plants, and the highest content was 0.850μg/g, which accounted for 0.016% of the total soluble protein, while no CrylAb protein was detected in some transgenic plants. There were also obvious differences in CrylAb protein content in the different organs of the same transgenic plants, following the order of leaf 〉 glumes 〉 seed 〉 root 〉 stem. The CrylAb protein content in middle leaf was higher than that in base and top leaf.
出处
《核农学报》
CAS
CSCD
北大核心
2009年第3期391-394,共4页
Journal of Nuclear Agricultural Sciences
基金
国家自然科学基金(30270273)
浙江省自然科学基金(Y304463)
