[苏云金芽孢杆菌变种tenebrionis的部分修饰δ-内毒素基因在转基因马铃薯植株中的表达]

[Expression of a partially modified delta-endotoxin gene from Bacillus thuringiensis var. tenebrionis in transgenic potato plants].

作者信息

Gulina I V, Shul'ga O A, Mironov M V, Revenkova E V, Kraev A S, Pozmogova G E, Iakovleva G A, Skriabin K G

出版信息

Mol Biol (Mosk). 1994 Sep-Oct;28(5):1166-75.

Abstract

A modified gene (Bt77) of delta-endotoxin from Bacillus thuringiensis var. tenebrionis was constructed and cloned into pMON505. This binary transformation vector was introduced into Agrobacterium tumefaciens strains containing different helper disarmed Ti-plasmids, LBA4404, A281, and CBE21. These Agrobacterium strains were used to transform potato stem segments (S. tuberosum, cv Desiree, Resy, Temp, Granat). Regenerants were selected on kanamycin-containing media. The presence of the Bt77 sequence in plant genomic DNA was confirmed by PCR analysis. Bt gene expression was studied in regenerated plants. Western blot analysis revealed that transgenic plants produced the Bt protein in the range of 0.005-0.02% of total protein. Total protection against insect damage of leaf tissue from these plants was observed in laboratory bioassays with of Colorado beetle larvae. Transgenic plants showed incomplete protection from CB larvae.

摘要

构建了苏云金芽孢杆菌变种暗黑变种δ-内毒素的修饰基因（Bt77），并将其克隆到pMON505中。将该二元转化载体导入含有不同辅助解除武装Ti质粒的根癌农杆菌菌株LBA4404、A281和CBE21中。这些根癌农杆菌菌株用于转化马铃薯茎段（S. tuberosum，品种Desiree、Resy、Temp、Granat）。在含卡那霉素的培养基上筛选再生植株。通过PCR分析证实了植物基因组DNA中Bt77序列的存在。对再生植株中的Bt基因表达进行了研究。蛋白质免疫印迹分析表明，转基因植物产生的Bt蛋白占总蛋白的0.005 - 0.02%。在科罗拉多马铃薯甲虫幼虫的实验室生物测定中，观察到这些植物的叶片组织对昆虫损伤具有完全保护作用。转基因植物对CB幼虫的保护作用不完全。

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