IL-13 cDNA探针的制备及应用

Preparation and Application of the IL-13 cDNA Probe

采用缺口平移的方法用生物素标记hIL-13的cDNA片段作为探针,检测传代培养的肿瘤细胞株中IL-13的基因表达,RNA斑点杂交和Northern杂交的结果均显示这些肿瘤细胞株有不同程度IL-13基因的表达.该探针本身的显色灵敏度较高,用5～10pg量的探针点膜可见明显的显色.探针的特异性良好,含IL-13mRNA的核酸杂交结果呈阳性,而不表达IL-13基因的RNA与该探针的杂交结果呈阴性.IL-13cDNA探针的制备为进一步研究IL-13的生物学特性提供有效的手段.

The gene expression of IL-13 on tumor cell lines were investigated with hIL-13 cDNA probe, which was labelled with biotin by nick translation. The results obtained by RNA dot blot and Northern blot showed that there were different degrees of IL-13 gene expressions on the tumor cells. The sensitivity of this probe was rather high with a visible colorations under 5~ 10pg of biotin-labelled probe dotted in NC membrane, and the specificity of the probe was also good. Ribonucleic acid (RNA) containing IL-13 mRNA hybridized with IL-13 cDNA probe and gave positive results, while RNA without IL-13 gene showed a negative result. The preparation of IL-13 probe lays foundation for further researches on the biological properties of IL-13.