Schreiber J H, Schisa J A, Wilson J M
University of Michigan.
Biotechniques. 1993 May;14(5):818-23.
Two histochemical marker genes, Drosophila alcohol dehydrogenase (ADH) and human placental alkaline phosphatase (ALP), were cloned into the recombinant retroviral vectors pLJ and pgag beta-actin. The resulting vectors were transfected into retroviral producer cell lines, psi CRE and psi CRIP, and stable recombinant retrovirus producers were isolated. Recombinant virus was harvested and used to transduce genes into several cell lines, singly or in conjunction with lacZ (Escherichia coli beta-galactosidase)-containing retrovirus. Cell lines were then stained using standard histochemical methods for recombinant gene expression. We found that multiple gene products could be identified in the same cell populations and in the case of ALP and beta-galactosidase, in the same cells. The resulting reagents should be useful for a variety of cell-marking studies including those involving multiple clonal analysis and developmental studies for gene therapy.
将两个组织化学标记基因，果蝇乙醇脱氢酶（ADH）和人胎盘碱性磷酸酶（ALP），克隆到重组逆转录病毒载体pLJ和pgagβ-肌动蛋白中。将所得载体转染到逆转录病毒生产细胞系psi CRE和psi CRIP中，并分离出稳定的重组逆转录病毒生产者。收获重组病毒并用于将基因单独或与含lacZ（大肠杆菌β-半乳糖苷酶）的逆转录病毒一起转导到几种细胞系中。然后使用标准组织化学方法对细胞系进行染色以检测重组基因表达。我们发现可以在相同的细胞群体中鉴定出多种基因产物，并且就ALP和β-半乳糖苷酶而言，可以在相同的细胞中鉴定出多种基因产物。所得试剂应可用于各种细胞标记研究，包括那些涉及多重克隆分析和基因治疗发育研究的研究。
