多药耐药基因Mdr1 RNA探针的构建及初步临床应用

检测Mdrl基因表达水平可预测白血病化疗效果，用原位杂交的方法可检测Mdrl在单个细胞的表达水平。本文用Rt-PCR的方法获得了一段特异的cDNA片段，将其克隆到PGEM4Z载体中，经DNA序列分析证明与文献报道一致，采用地高辛素（DIG）RNA标记试剂盒制备反义RNA探针，已初步用于临床骨髓涂片标本的原位杂交检测。

CONSTRUCTION AND PRIMARY APPLICATION OF Mdrl GENE RNA PROBE

The chemotherapy effects of patients with leukemia are correlated with the expression level of Mdrl gene which can be measured by ISH in individual leukemic cells because of the cDNA cloning of the human Mdrl. A special part of Mdrl cDNA was obtained by Rt PCR method and inserted into vector PGEM4Z. The DNA sequencing result showed it was identical with the sequence of Mdrl cDNA reported. The Mdrl RNA probe was made from the cDNA by use of DIG RNA labeling kit. The detecting system of Anti Dig AP was used on leukemia marrow smears and showed high specificity and sensibility.