人细胞差异表达基因cDNA消减杂交体系的建立

A Strategy for Isolating Differentially Expressed Gene From Human Cells.

为克隆肺腺癌分化相关基因，采用诱导分化与消减杂交相结合的策略，建立了全反式维甲酸（ＲＡ）诱导前后人肺腺癌细胞系的ｃＤＮＡ消减文库，得到１２４个ｃＤＮＡ消减克隆．经加减法杂交差异筛选、ＤＮＡ和ＲＮＡ印迹、ｃＤＮＡ全序列测定和生物学功能分析，分离到３个在人肺腺癌细胞系分化过程中由ＲＡ激活而特异表达的新的ｃＤＮＡ序列这一策略和技术路线适用于分离细胞中呈过量表达或表达抑制基因的ｃＤＮＡ克隆，并具有反映细胞分化过程中基因表达动态变化特征和相对简便适用的特点．

A subtractive hybridization system for isolating differentiation associated gene from human cancer cell line is developed. By using the strategy of differentiation-inducing in combina tion with cDNA-cDNA subtractive hybridization, a subtracted cDNA library from human lung adenocarcinoma cell line before and after treatment with all-trans retinoic acid(RA) was established. Three cDNAs response to RA treatment were obtained by screening of the subtracted cDNA library. The results of sequencing and function analysis show that three cDNAs are novel cDNAs of differentiation associated genes in human lung adenocarcinoma cell line. This system is also suitable for isolating cDNAs representing deleted or overexpressed gene in human cells.