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７３５ｂｐ的ＰＲＬｃＤＮＡ片段从质粒ＰＲＬ－ＳＰ６５＃１中回收后，用粘性末端连接法将其重组到真核表达载体ｐｃＤＮＡ３上，筛选出正向连接重组体ｐｃＤＮＡ３－ＰＲＬＳ和反向连接重组体ｐｃＤＮＡ３－ＰＲＬＡＳ。将重组体ｐｃＤＮＡ３－ＰＲＬｓ和空载体ｐｃＤＮＡ３分别转入ＮＩＨ３Ｔ３细胞系，用Ｇ４１８筛选出阳性细胞后与未转染的ＮＩＨ３Ｔ３细胞在加Ｅ２和不加Ｅ２的情况下，用原位杂交的方法，分别用ＰＲＬｃＤＮＡ探针和原癌基因ｃ－Ｈ－ｒａｓｃＤＮＡ探针进行检测，未转染的ＮＩＨ３Ｔ３细胞在加Ｅ２和不加Ｅ２时都几乎无催乳素基因的表达，同样，转入空载体的ＮＩＨ３Ｔ３细胞也无ＰＲＬ的表达，而转入重组体ｐｃＤＮＡ３－ＰＲＬＳ的ＮＩＨ３Ｔ３细胞则有大量的ＰＲＬ基因的表达，与对照组相比有显著差异（Ｐ＜０．０１）。正常和转入空载体的ＮＩＨ３Ｔ３细胞有一定程度的原癌基因ｃ－Ｈ－ｒａｓ的表达，当分别加入Ｅ２和转入重组体ｐｃＤＮＡ３－ＰＲＬＳ后，ＮＩＨ３Ｔ３细胞中的ｃ－Ｈ－ｒａｓ基因表达水平都显著升高（Ｐ＜０．０５）。
The PRL cDNA fragment of 735bp was recovered from the plasmid PRL-SP65 # 1 and then recombined into the eukaryotic expression vector pcDNA3 by the sticky end connection method. The positive recombinant pcDNA3-PRLS and the reverse recombinant pcDNA3-PRLAS . The recombinant pcDNA3-PRLs and empty vector pcDNA3 were transferred to NIH3T3 cell line, positive cells were screened with G418 and untransfected NIH3T3 cells in the case of adding E2 and without E2, using in situ hybridization method, respectively Using PRLcDNA probe and proto-oncogene c-H-rascDNA probe, untransfected NIH3T3 cells showed almost no expression of prolactin gene with and without E2, and transfected into empty vector NIH3T3 cells There was also no expression of PRL, while NIH3T3 cells transfected with recombinant pcDNA3-PRLS had a large number of PRL gene expression compared with the control group (P <0.01). NIH3T3 cells in normal and transfected NIH3T3 cells had a certain degree of expression of c-H-ras, and the expression of c-H-ras gene in NIH3T3 cells after addition of E2 and recombinant pcDNA3-PRLS respectively Were significantly higher (P <0.05).