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Construction of ATP7B gene recombinant adenovirus vector Ad-ATP7B in Wilson's disease
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- Author:
- No author available
- Journal Title:
- CHINESE JOURNAL OF NEUROMEDICINE
- Issue:
- 6
- DOI:
- 10.3760/cma.j.issn.1671-8925.2005.06.001
- Key Word:
- 肝豆状核变性;腺病毒载体;基因疗法
Abstract： 目的构建Wilson病基因ATP7B的重组腺病毒载体.方法分别以BamH Ⅰ+SalⅠ双酶切pcDNA3.0/ATP7B和pDC315,将ATP7B cDNA目的基因片段和线性化的pDC315连接,定向克隆构建pDC315/ATP7B,以PCR和酶切的方法鉴定.pDC315/ATP7B与腺病毒骨架共转染293细胞构建Ad-ATP7B,PCR进行鉴定. 结果经PCR和酶切鉴定证实pDC315/ATP7B构建成功.pDC315/ATP7B与腺病毒骨架共转染293细胞后见明显的毒斑,说明二者在293细胞中同源重组并包装成功.经PCR证实重组腺病毒Ad-ATP7B构建已完成.结论本实验成功构建了ATP7B外源目的基因序列完全正确的重组腺病毒载体Ad-ATP7B,为下一步采用ATP7B基因重组腺病毒载体对Wilson病进行基因治疗打下了基础.
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