人牙周膜成纤维细胞与牙龈成纤维细胞差异表达基因扣除文库的构建

摘要

AIM: To construct a subtractive cDNA library of genes differentially expressed in cultured primary human PDLF in comparison with GF. METHODS: mRNAs were isolated fiom primary cultures of human PDLF and GF. Subtractive eDNA library of PDLF was constructed with a recently developed gene cloning technique that is based on PCR and subtractive hybridization. Part of randomly selected clones were identified by restriction analysis and reverse Southern dotblot. RESULTS: A subtbtetive cDNA library of PDLF was constructed with a capieity of 4×l02 clones. CONCLUSION:The gene cloning method we used in this study is both simple and effective in cloning differentially expressed gene.%目的：建立人牙周膜成纤维细胞(periodontal ligament fibroblast, PDLF)rn与牙龈成纤维细胞(gin-gival fibroblast, GF)差异表达基因的扣除文库。方法：体外培养人PDLC和GF，分别提取mRNA，采用基于PCR和消减杂交的基因克隆技术构建人PDLC与GF差异表达基因的扣除文库。结果：成功构建了人PDLF与GF细胞差异表达基因的扣除文库，文库容量为4×l02。结论：基于PCR和消减杂交的基因克隆技术是构建细胞间差异表达基因扣除文库的较为简洁而有效的方法。