摘要
目的构建GDNF真核表达载体,为进一步治疗脊髓损伤打下基础。方法首先提取新生大鼠肾组织总RNA,用逆转录PCR法获得并扩增GDNF全长cDNA,构建其真核表达载体pcDNA3.1/GDNF,双酶切电泳鉴定,并进行序列测定。结果扩增到全长GDNF cDNA,构建载体测序及酶切鉴定证实克隆成功。结论成功构建带有真核启动子的GDNF真核表达载体,为下一步转基因应用到脊髓损伤后的治疗奠定了基础。
[ Objective ] To explore the role of glial cell-line derived neurotrophic factor （GDNF） in the treatment of SCI in rats and construct eukaryotic vector. [ Methods ] Extract total RNA from rat kidney firstly, and obtain GDNF eDNA by RT-PCR, and construct vector of pcDNA3.1/GDNF identified by double enzyme digestion and sequence analysis. [Result] Total DNA sequence of GDNF was obtained, which was demonstrated by double enzyme digested and sequence analysis. [Conclusion] GDNF eukaryotic vector has been constructed successfully, which lays the foundation for the next transgene study of the treatment of spinal cord injury.
出处
《中国医学工程》
2009年第3期167-170,共4页
China Medical Engineering
