差异筛选肝癌凋亡细胞cDNA文库的简便方法

目的差异筛选人肝癌凋亡细胞ｃＤＮＡ文库。方法消减杂交和点杂交相结合的噬菌斑原位杂交法筛选ｃＤＮＡ文库，首先采用（－）ｃＤＮＡ探针杂交，挑取（－）的噬菌斑克隆，再用（－）和（＋）两种ｃＤＮＡ探针与初筛出的噬菌斑克隆杂交的差异筛选方法。结果得到４个充分孤立的噬菌斑克隆，其插入片段长度为１．５ｋｂ左右。结论该方法简便、快速，是差异筛选ｃＤＮＡ文库的一种较为可行的简便方法。

A RAPID METHOD FOR SUBTRACTIVE SCREENING OF HEPATOMA APOPTOTIC CELLS cDAN LIBRARY

Objective To expedite the subtractive screening of human hepatoma apoptotic cells cDNA library.Methods The method of subtractive hybridization combined with dot blot hybridization was adopted. First, minus cDNA probe was used to screen the cDNA library, the minus clones that did not hybridize with the minus cDNA probe were picked up as the source of the second grade dot blot hybridization. Secondly, two probes of the plus and minus cDNA were used to screen the clones; the clones that only hybridize with the plus cDNA probe were picked up as the source of the third grade dot blot hybridization.Results Four clones were obtained and the lengths of the inserted cDNA fragments were about 1.5kb long. Conclusion The results demonstrate this is a feasible, simple and quick method for subtractive screening of the cDNA library.