医学分子生物学杂志2012，Vol.9Issue(5)：355-359,5.DOI:10.3870/j.issn.1672-8009.2012.h5.008
分泌型碱性磷酸酶报告基因重组载体pSEAP-IQCE3'UTR的构建
Construction of Recombinant Plasmid pSEAP-IQCE3'UTR
摘要
Abstract
Objective To construct the recombinant vector pSEAP-IQCE3'UTR containing secreted alkaline phosphatase reporter gene and explore whether FXR1P plays its role by interacting with IQCE3'UTR. Methods The PCR primers for the target gene fragment of IQCE3'UTR were designed, and then the IQCE3'UTR gene fragments were amplified by RT-PCR. The restriction endo-nuclease Xba Ⅰ was used for digestion of the amplified fragments and the reporter gene empty vector. The empty vectors were dephosphorylated and then connected with the target fragment. The resulting product was transformed into E. coli JM109, and the positive clones were selected. The sequence was detected by PCR. Results Restriction endonuclease digestion, positive and negative i-dentification and sequencing analysis showed that the recombinant vector pSEAP-IQCE3'UTR containing secreted alkaline phosphatase reporter gene was successfully constructed. Conclusion The successfully constructed recombinant vector pSEAP-IQCE3'UTR is of great significance in further exploring the function of FXR1P and its pathogenesis in fragile X syndrome.关键词
FXR1P/IQCE/SEAP/脆性X综合征Key words
FXR1P/ IQCE/ secreted alkaline phosphatase reporter gene (SEAP) / fragile X syndrome分类
生物科学引用本文复制引用
王三虎,马云,秦凌雪,何彬彬,李斌元,苏泽红,何淑雅..分泌型碱性磷酸酶报告基因重组载体pSEAP-IQCE3'UTR的构建[J].医学分子生物学杂志,2012,9(5):355-359,5.基金项目
湖南省自然科学基金(No.12JJ6073) （No.12JJ6073）
