目的制备用地高辛 (digoxigenin ,Dig)标记的血小板T细胞活化抗原1(plateletandTcellactivationantigen1,PTA1)cRNA探针。方法构建重组质粒 pGEM 3ZF( +) PTA1 ,并做序列分析证实PTA1基因的插入方向正确后 ,用EcoRI消化得到线性DNA片段 ,再用SP6RNA聚合酶转录合成带有Dig标记的高比活度的单链cRNA探针。结果经斑点杂交证实 ,该探针敏感性高、特异性强。结论地高辛标记PTA1cRNA探针的制备 ,为进一步研究PTA1mR NA在组织、细胞的表达和分布提供了有效的工具。 Objective To prepare digoxigenin (Dig) labeled cRNA probe for platelet T cell activation antigen 1 (PTA1). Methods The recombinant plasmid pGEM 3ZF (+) PTA1 was constructed and sequenced to confirm the correct insertion direction of PTA1 gene. After digestion with EcoRI, a linear DNA fragment was obtained and then transcribed with SP6 RNA polymerase to generate a single tag with high specific activity Chain cRNA probe. The results confirmed by dot blot hybridization, the probe sensitivity and specificity. Conclusion The preparation of digoxigenin-labeled PTA1cRNA probe provides an effective tool for further study on the expression and distribution of PTA1mRNA in tissues and cells.