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本实验采用含生长抑素基因的PSP65cDNA质粒通过转化至大肠杆菌扩增,经碱变性提取,纯化后用限制性内切酶进行酶切使质粒线性化,并将其作为模板,用Digoxigenin-UTP作为标记物(Boehringer公司)体外转录合成生长抑素(Somatostatin,SS)的反意 RNA探针。选用新生大鼠经多聚甲醛心脏灌注固定后,速取全脑,恒冷箱冰冻切片,切片经杂交前预处理后用含有Digoxigenin-UTP标记的SS反意 RNA探针杂交液进行杂交,之后切片,经后处理予以显色。结果表明含有生长抑素
In this experiment, the plasmid containing somatostatin gene of PSP65cDNA was amplified by transformation into E.coli. The plasmid was extracted by alkaline denature and purified by restriction endonuclease digestion to linearize the plasmid, which was then used as a template. Digoxigenin-UTP In vitro transcript synthesis of somatostatin (SS) anti-sense RNA probe as a marker (Boehringer). Neonatal rats were selected and fixed with paraformaldehyde for cardiac perfusion. The whole brain and frozen cubes were cryopreserved in frozen flasks. The hybridized pre-hybridized sections were stained with Digoxigenin-UTP-labeled SS antisense RNA probe and then hybridized. Slice, after treatment to be color. The results show that somatostatin is contained