苏云金芽胞杆菌cry26Aa和cry28Aa基因共表达可在芽胞外壁内侧形成伴胞晶体

Co-expression of crystal protein gene cry26Aa and cry28Aa has an ability to form parasporal crystal inside exosporium in Bacillus thuringiensis subsp. finitimus

苏云金芽胞杆菌幕虫亚种的伴胞晶体在预芽胞外壁内侧形成,呈现晶体芽胞粘连的现象。根据已发表的cry26Aa1和cry28Aa1基因序列设计引物,从苏云金芽胞杆菌幕虫亚种T02中扩增得到cry26Aa和cry28Aa基因,通过穿梭载体将这两个基因分别和同时转化到苏云金芽胞杆菌无晶体突变株BMB171后,透射电镜下可在芽胞外壁内侧和外侧同时观察到伴胞晶体,而单独表达时可在芽胞外壁外侧观察到伴胞晶体。结果表明,伴胞晶体在芽胞外壁内侧表达不单独依赖于启动子的时空调控,可能还受到晶体蛋白相互作用的影响。

Parasporal crystal in Bacillus thuringiensis subsp, finitimusT02 forms within the exosporium and remains attached to the spore after mother cell lysis, which leads to spore-crystal connection. According to the crystal protein gene sequence in B. thuringiensis subsp, finitimus, cry26 and cry28 were cloned from strain T02 by PCR amplification and transformed into the crystal negative Bacillus thuringiensis strain BMB171 by shuttle vectors, alone or in combination together. Crystal can be observed inside of exosporium when co-expression of these two genes. But crystal mainly formed outside of exosporium when they expressed alone. This suggests interaction between of the two crystal proteins may be involved in enclosure.