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Construction of a DNA vaccine based on the MG7-Ag mimotope of gastric cancer
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- Author:
- No author available
- Journal Title:
- JOURNAL OF THE FOURTH MILITARY MEDICAL UNIVERSITY
- Issue:
- 2
- DOI:
- 10.3321/j.issn:1000-2790.2002.02.004
- Key Word:
- 胃肿瘤;分子模拟;表位;疫苗,DNA
Abstract： 目的构建以胃癌MG7-Ag模拟表位为基础的DNA疫苗. 方法将HindⅢ酶切位点、 MG7-Ag模拟表位和通用性辅助性T细胞(Th细胞)表位编码序列的反向互补序列顺次设计于下游引物中,通过2次聚合酶链式反应将2种表位连接于一段载体基因,HindⅢ酶切位点位于载体片段与表位基因之间. 将目的基因插入pUCm-T载体,酶切鉴定和序列测定证实后,利用 pUCm-T载体和pcDNA3.1(+)载体共有的KpnⅠ和XbaⅠ酶切位点,将目的基因亚克隆入真核表达载体pcDNA3.1(+)载体. 酶切鉴定后,利用目的基因和pcDNA3.1(+)载体上的HindⅢ酶切位点, HindⅢ酶切去除载体基因片段,得到胃癌MG7-Ag模拟表位和通用性Th细胞表位的真核表达载体. 结果经序列测定证实:PCR产物为载体基因、HindⅢ酶切位点、MG7-Ag模拟表位及通用性Th细胞表位的融合片段;最终得到的重组pcDNA3.1(+)质粒含有正确编码以上2种表位的基因片段. 结论以胃癌MG7-Ag模拟表位为基础的DNA疫苗构建成功 ,为进一步研究其在胃癌免疫治疗中的作用奠定了基础.
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