Ravoahangimalala O, Charles J F
Bactéries Entomopathogènes, Institut Pasteur, Paris, France.
FEBS Lett. 1995 Apr 3;362(2):111-5. doi: 10.1016/0014-5793(95)00220-4.
Midguts from Anopheles gambiae fourth instars were dissected and processed for immuno-light microscopy. Cloned insecticidal crystal proteins (ICPs) from Bacillus thuringiensis var. israelensis (Bti) were individually expressed in crystal-negative strains of Bacillus thuringiensis. Tissue sections of A. gambiae were incubated in vitro with each solubilized and trypsin-activated ICP. Immunodetection of CryIVA, CryIVB, CryIVD and CytA toxins on sections was performed using purified rabbit IgG directed against Bti ICPs, in combination with an anti-rabbit IgG/peroxidase. CryIVA, CryIVB, CryIVD and CytA toxins were detected on the apical brush border of midgut cells, in the gastric caecae and posterior stomach. CytA was also detected, to a lesser extent, on microvilli of anterior stomach cells.
解剖冈比亚按蚊四龄幼虫的中肠并进行处理，用于免疫光学显微镜观察。从苏云金芽孢杆菌以色列变种（Bti）克隆的杀虫晶体蛋白（ICPs）分别在苏云金芽孢杆菌的晶体阴性菌株中表达。将冈比亚按蚊的组织切片与每种溶解并经胰蛋白酶激活的ICP在体外孵育。使用针对Bti ICPs的纯化兔IgG结合抗兔IgG/过氧化物酶，对切片上的CryIVA、CryIVB、CryIVD和CytA毒素进行免疫检测。在中肠细胞的顶端刷状缘、胃盲囊和后胃中检测到CryIVA、CryIVB、CryIVD和CytA毒素。在胃前细胞的微绒毛上也检测到了CytA，但程度较轻。
