Expression of human multidrug resistance genes and improved selection of cells transduced with such genes

摘要

A DNA sequence for a human mdr1 gene, which encodes p- glycoprotein, wherein at least one base in a splice region of the DNA encoding p- glycoprotein is changed. Such a mutation prevents truncation of the p- glycoprotein upon expression thereof. There is also provided a method of identifying cells which express the human mdr1 gene in a cell population that has been transduced with an expression vehicle including a human mdr1 gene. The method comprises contacting the cell population with a staining material, such as rhodamine 123, and identifying cells which express the human mdr1 gene based on differentiation in color among the cells of the cell population. This method has allowed identification of retroviral producer clones facilitate mdr gene transfer into primary cells. Repopulating hematopoietic stem cells have been genetically engineered with the human mdr1 gene.