东亚钳蝎抑制型神经毒素基因在昆虫细胞中的表达及活性鉴定

BACULOVIUS-MEDIATED EXPRESSION OFA SCORPION DEPRESSANT TOXIN AND BIOACTIVITY ASSAY

基因BmKIT3R 是根据东亚钳蝎 (ButhusmartensiiKarsch)抑制型神经毒素基因BmKIT3 优化而得的 .将优化过的蝎毒素基因BmKIT3R 通过Bac to Bac操作技术 ,使重组杆状病毒基因组DNA转染到草地粘虫sf2 1细胞中对IT3R 基因进行表达 ,经SDS PAGE检测在 8.7× 10 3 处有一表达条带 .表达产物经生物鉴定 ,证明具有很高的活性 ,它对昆虫具有非常明显的致死作用 .图 3参

The optimized Buthus martensii Karsch gene was ligated into the pFastBac1 donor plasmid which had been linear zed and then transformed into E.coli DH10 competent cells. Transformed colonies containing the recombinant bacmid were identified by blue-white screening. Finally, bacmid DNA containing the BmK IT 3R cDNA insert was used to transfect sf21 armyworm ovary cell. SDS-PAGE analysis displayed a strong band at the position around 8.7×103. The activity of the toxins produced by the recombinant baculovirus was assessed by injection of 5 th silkworm larvae. Fig 3, Ref 5