人IL—3cDNA探针制备及应用

Preparation and application of human IL - 3cDNA probe

采用多聚酶连反应（ＰＣＲ）扩增人ＩＬ—３ｃＤＮＡ片段，以地高辛（Ｄｉｇｏｘｉｇｅｎｉｎ—１１—ｄＵＴＰ）为标记物，随机引物法标记ＩＬ—３ｃＤＮＡ探针。探针经斑点杂交和核酸原位杂交证明：用制备ＩＬ—３ｃＤＮＡ探针检测细胞ＩＬ－－３ｍＲＮＡ表达具有特异性强，灵敏度高，使用简便，杂交结果直观，探针可较长时间保存和无放射性污染等优点。

By using the techniques of polymerase chain reaction(PCR),human IL - 3cDNA probe was prepared with pUC18- IL- 3 plasmid, labeled with digoxigenin - 11dUTP and identified with dot blot. We detected the expressionof IL- 3 mRNA in murine bone marrow stromal cells, human fetal thymocytes and splenic cells by nucleisitu hybridization. The results indicated that the prepared IL-3cDNA probe can provide the sensitivitysimplicity of handling needed to replace radioactiassays.