Tec家族蛋白酪氨酸激酶Itk特异地表达于T细胞 ,对T细胞的成熟与活化等具有很大影响 ,因而可能在T细胞的信号转导过程中处于重要位置。我们利用PCR的方法从含Itk基因全长cDNA的pME18S EMT质粒中扩增出其PH结构域基因片段 ,并将其克隆入酵母双杂交载体pLexA ,经测序鉴定正确 ,转化酵母细胞EGY48(p8op lacZ ) ,挑取阳性克隆 ,利用液相法和平板法检测 ,证实PH结构域无自激活作用 ,且对宿主菌无毒性作用 ,可用于T细胞cDNA文库的筛选 ,捕捉与之相互作用的蛋白。 Itk family protein tyrosine kinase Itk is specifically expressed in T cells and has a great influence on the maturation and activation of T cells, which may play an important role in the signal transduction of T cells. We amplified the PH gene fragment from the pME18S EMT plasmid containing the full-length cDNA of Itk gene by PCR and cloned it into the yeast two-hybrid vector pLexA. The recombinant plasmid was identified by sequencing and transformed into yeast cell line EGY48 (p8op lacZ ). The positive clones were picked and tested by liquid phase and plate method. The results showed that PH domain had no self-activation and had no toxic effect on the host bacteria. It could be used to screen T cell cDNA library and capture the protein interacting with it.