论文部分内容阅读
以成年ＳＤ大鼠坐骨神经损伤后的远段为材料，以λｇｔ１１为载体，构建了ＳＤ大鼠损伤性坐骨神经的ｃＤ－ＮＡ基因文库。将文库以５×１０４／１２０ｍｍ培养皿的密度铺皿培养，８ｈ后将印迹的硝酸纤维素膜与特异性６５ＫＤ单克隆抗体混合物４℃孵育过夜。行免疫组化ＡＢＣ反应，筛选４×１０５克隆后得到３个阳性信号点。挑出相应的克隆，进行第２、３轮筛选，得到表达６５ＫＤ化学诱向因子的单克隆。该实验为基因水平研究周围神经再生发育提供依据
The cD-NA gene library of injured rat sciatic nerve in SD rats was constructed by using λgt11 as a vector in the distal part of sciatic nerve of adult SD rats. Libraries were plated at a density of 5 x 104/120 mm dishes and after 8 h the blotted nitrocellulose membranes were incubated with a specific 65 kD monoclonal antibody mixture overnight at 4 ° C. ABC immunohistochemical reaction, screening 4 × 105 clones obtained 3 positive signal points. The corresponding clones were picked out and the second and the third rounds of screening were carried out to obtain a single clone expressing the chemotactic factor of 65KD. This experiment provides the basis for the study of peripheral nerve regeneration and development at the gene level