携带NR2B抗原表位的表达载体的构建与鉴定

目的构建携带N-甲基-D-天门冬氨酸受体(N-methyl-D-aspartate receptor,NMDAR)2B亚基抗原表位的表达载体并进行初步鉴定。方法在获得NMDAR2B(NR2B)抗原模拟表位碱基序列TCGCATCCGCCTGTGATGCCTTGGCC-TACTTCGACT的基础上,设计两条含起始密码子、终止密码子、双酶切位点的互补引物,应用PCR技术合成该表位,与表达载体pDC515重组为pDC515/nr2b,采用限制性内切酶酶切及DNA测序进行鉴定。结果经限制性酶切鉴定及DNA测序分析证实pDC515/nr2b含有NR2B抗原模拟表位片段。结论成功构建了含有NR2B模拟抗原表位的表达载体,为进一步研制NR2B抗原表位的DNA疫苗及其镇痛效应奠定了实验基础。

Construction and identification of plasmid vector with NR2B mimic epitope

Objective To construct and identify a recombinant plasmid vector with NR2B mimic epitope. Methods The DNA of specific phage clone expressing a mimic epitope of NR2B was purified and sequenced.The sequence was TCGCATCCGCCTGTGATGCCTTGGCCTACTTCGACT.The target fragment of NR2B mimic epitope was obtained by PCR,then recombined with the expression vector pDC515 and identified by PCR,restricted endonuclease and DNA sequence analysis.Results The restriction endonuclease digestion and DNA sequencing confirmed that the recombined plasmid vector pDC515/nr2b contained a mimic epitope of NR2B.Conclusion A recombined plasmid vector pDC515/nr2b with a mimic epitope of NR2B has been successfully constructed.The experiment will establish the basis for studying the DNA vaccine of NR2B and gene analgesia.