Yan Hong, Kong Dong, Ge Xiaomei, Gao Xiang, Han Xiao
Key Laboratory of Human Functional Genomics of Jiangsu Province, Nanjing Medical University, Nanjing, Jiangsu 210029, China; ; Model Animal Research Center, Nanjing University, Nanjing, Jiangsu 210093, China.
J Biomed Res. 2011 Nov;25(6):438-43. doi: 10.1016/S1674-8301(11)60058-4.
Phosphatase of regenerating liver-3 (PRL-3) is a member of the protein tyrosine phosphatase (PTP) superfamily and is highly expressed in cancer metastases. For better understanding of the role of PRL-3 in tumor metastasis, we applied a rapid and efficient method for generating PRL-3 floxed mice and investigated its phenotypes. A BAC retrieval strategy was applied to construct the PRL-3 conditional gene-targeting vector. Exon 4 was selected for deletion to generate a nonfunctional prematurely terminated short peptide as it will cause a frame-shift mutation. Conditional knockout PRL-3 mice were generated by using the Cre-loxP system and were validated by Southern blot and RT-PCR analysis. Further analysis revealed the phenotype characteristics of PRL-3 knockout mice and wildtype mice. In this study, we successfully constructed the PRL-3 conditional knockout mice, which will be helpful to clarify the roles of PRL-3 and the mechanisms in tumor metastasis.
再生肝脏磷酸酶-3（PRL-3）是蛋白质酪氨酸磷酸酶（PTP）超家族的成员，在癌症转移中高表达。为了更好地理解PRL-3在肿瘤转移中的作用，我们应用了一种快速有效的方法来生成PRL-3基因条件性敲除小鼠并研究其表型。采用BAC检索策略构建PRL-3条件性基因打靶载体。选择外显子4进行缺失，以产生无功能的过早终止短肽，因为这会导致移码突变。利用Cre-loxP系统产生条件性敲除PRL-3的小鼠，并通过Southern印迹和RT-PCR分析进行验证。进一步分析揭示了PRL-3敲除小鼠和野生型小鼠的表型特征。在本研究中，我们成功构建了PRL-3条件性敲除小鼠，这将有助于阐明PRL-3在肿瘤转移中的作用及其机制。
