牙根发育不全疾病致病差异表达基因文库的建立

目的：建立先天性牙根发育不全疾病患儿与其正常兄弟间致病差异表达基因的cDNA文库。方法：采用改良消减杂交技术，以先天性牙根发育不全疾病患儿与其正常兄弟外周静脉血的总mRNA为对比材料，筛选出候选相关致病基因差异表达的cDNA，将其与pGEM-T载体进行T／A连接构建文库，然后转化高效感受态大肠杆菌进行文库扩增，随机挑取100个白色克隆进行酶切鉴定。结果：扩增消减cDNA文库获得400余个白色阳性克隆，随机挑取的100个白色克隆经酶切后90％有插入片段。结论：用改良消减杂交技术成功构建了先天性牙根发育不全疾病致病差异表达基因的消减cDNA文库，该文库的建立为进一步筛选、克隆该疾病的候选致病相关基因奠定了基础。

Construction of Cdna library subtracted from congenital hypoplasia of teeth roots

Objective:To construct the differentially expressed cDNA library subtracted from the patient of congenital hypoplasia of teeth roots and normal brother. Method:By modified subtractive hybridization technique, total RNA sample of Peripheral venous blood extracted from the patient of congenital hypoplasia of teeth roots and normal brother were subtracted to screen the differentially expressed cDNA of candidate virulence gene related with this disease. Then the cDNA were ligated with pGEM-T vector, and the inserted vector were transformed to competence Escherichia coli for library amplification.100 white clone were selected randomly and evaluate by enzyme digestion.Result:400 white cloning were obtained in the cDNA library, and after enzyme digestion there were inserted fragmentsin 90% of 100 cloning selected randomly.ConclusionThe differentially expressed cDNA library was constructed successfully, which was based on screening and cloning candidate virulence gene related with this disease.