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Cloning and identification of human full-length TSLC1 cDNA and construction of its eukaryotic expression vector
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- Author:
- No author available
- Journal Title:
- JOURNAL OF THE FOURTH MILITARY MEDICAL UNIVERSITY
- Issue:
- 10
- DOI:
- 10.3321/j.issn:1000-2790.2007.10.003
- Key Word:
- TSLC1基因;pcDNA3.1(+);克隆,分子
Abstract： 目的:克隆人类TSLC1基因并构建其真核表达载体pcDNA3.1(+)/TSLC1.方法:从正常皮肤组织中提取总RNA,采用RT-PCR方法扩增TSLC1基因全长cDNA,克隆入pMD18-T载体并转化大肠杆菌JM109,经PCR、酶切鉴定均为阳性的克隆,进行核苷酸测序分析,再将TSLC1基因定向克隆入pcDNA3.1(+)载体中构建表达载体.结果:RT-PCR扩增后的产物在约1413 bp处出现明显的特异性条带,TSLC1基因的cDNA片段被成功插入真核表达载体pcDNA3.1(+)质粒的多克隆位点,经鉴定与GenBank收录的TSLC1 cDNA 序列一致.结论:TSLC1基因的cDNA片段被成功克隆,为以后的研究奠定了基础.
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