摘要
目的:构建pBiFC-VN173-Olig1和pBiFC-VC155-Id2真核表达质粒,并进行鉴定。方法:以pEGFP-N3-Olig1真核表达质粒为模板扩增出Olig1基因,与pBiFC-VN173载体连接,构建pBiFC-VN173-Olig1真核表达质粒;利用RT-PCR方法从新生大鼠脊髓中提取Id2基因片段,与pBiFC-VC155载体连接,构建pBiFC-VC155-Id2真核表达质粒。对此2种质粒进行酶切鉴定、测序。结果:通过酶切鉴定、测序,证明pBiFC-VN173-Olig1和pBiFC-VC155-Id2重组质粒序列和编码框均正确构建成功。结论:成功构建了pBiFC-VN173-Olig1和pBiFC-VC155-Id2真核表达质粒,为进一步在活细胞内研究Olig1和Id2的相互作用提供了实验基础。
Objective：To construct and identify pBiFC-VN173-Olig1 and pBiFC-VC155-Id2 eukaryotic expression plasmid for bimolecular fluorescence complementation（BiFC）assay.Methods：Rat olig1 gene from pGFP-N3-Olig1 eukaryotic expression plasmid was amplified by PCR.And rat Id2 gene from RNA of neonatal rat spinal cord was amplified by RT-PCR.The Olig1 gene was inserted into BiFC eukaryotic expression vector pBiFC-VN173 and Id2 gene was inserted into pBiFC-VC155,which were used to construct recombinant expression vector pBiFC-VN173-Olig1 and pBiFC-VC155-Id2,respectively.The recombinant vectors were identified by restriction enzyme digestion and DNA sequencing.Results：The restriction enzyme digestion and DNA sequencing results showed that the sequences and open read frames of the two vectors were completely concordant with experiment design.Conclusions：The pBiFC-VN173-Olig1 and pBiFC-VC155-Id2 were successfully constructed.These provide an experimental base for further research on interaction between Olig1 and Id2 in vivo.
出处
《蚌埠医学院学报》
CAS
2010年第10期973-975,共3页
Journal of Bengbu Medical College
基金
国家自然科学基金资助项目(30700439
81071268)
教育部科学技术研究重点资助项目(210103)
安徽省第五批(2010年度)优秀青年科技基金资助项目(10040606Y13)
安徽省教育厅自然科学研究项目(KJ2010B109)
关键词
基因
Olig1
ID2
克隆
酶切
gene
Olig1
Id2
cloning
restriction enzyme digestion
作者简介
郭术俊（1977-），女，实验师．
[通讯作者]胡建国，男，博士，研究生导师，副教授，E-mail：jghu．hzlv@yahoo．com
