CAJ | 학술논문

目的：原核表达人Pin2/TRF1结合蛋白X1（PinX1），确定该蛋白与人端粒酶逆转录酶（hTERT）催化亚基的相互作用。方法：用PCR方法从乳腺文库中扩增PinX1基因的编码序列，克隆至pET-32a载体构建重组质粒pHis-PinX1，经双酶切鉴定后转化大肠杆菌BL21并进行诱导表达，SDS-PAGE和Western印迹检测His-PinX1的表达；用His磁珠纯化His-PinX1，在人肾胚细胞293T内检测His-PinX1与hTERT的相互作用。结果：扩增得到980 bp的PinX1基因；Western印迹检测表明，相对分子质量约57×103的His-PinX1获得表达，且纯化的His-PinX1与hTERT具有相互作用。结论：表达并纯化得到了与hTERT相互作用的His-PinX1，为深入研究PinX1的功能奠定了基础。
목적：원핵표체인Pin2/TRF1결합단백X1（PinX1），학정해단백여인단립매역전록매（hTERT）최화아기적상호작용。방법：용PCR방법종유선문고중확증PinX1기인적편마서렬，극륭지pET-32a재체구건중조질립pHis-PinX1，경쌍매절감정후전화대장간균BL21병진행유도표체，SDS-PAGE화Western인적검측His-PinX1적표체；용His자주순화His-PinX1，재인신배세포293T내검측His-PinX1여hTERT적상호작용。결과：확증득도980 bp적PinX1기인；Western인적검측표명，상대분자질량약57×103적His-PinX1획득표체，차순화적His-PinX1여hTERT구유상호작용。결론：표체병순화득도료여hTERT상호작용적His-PinX1，위심입연구PinX1적공능전정료기출。
Objective: To express Pin2/TRF1-interacting protein X1(PinX1) in E.coli, and verify interaction be-tween purified His-PinX1 and human telomerase reverse transcriptase(hTERT). Methods: The coding sequence of PinX1 was amplified by PCR and fused in frame with the coding region of the pET-32a vector to generate pHis-PinX1. His-PinX1 was expressed in E.coli BL21 and identified by SDS-PAGE and Western blotting. The interac-tion between purified PinX1 and hTERT was detected in 293T cell. Results: The 980 bp PinX1 gene was insert-ed into pET-32a vector, and its expression product with relative molecular weight 57×103 was identified by SDS-PAGE and Western blotting. Purified His-PinX1 interacted with hTERT in 293T cell. Conclusion: The recombi-nant PinX1 which can interact with hTERT has been obtained, laying a foundation for further study on the func-tion of PinX1.