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目的：利用基因重组方法构建出含有抑癌基因 Ｐ１６， Ｐ５３的融合表达载体。方法：用限制性内切酶 Ｅｃｏ Ｒ Ｉ、 Ｘｈｏ Ｉ双酶切 Ｐ１６ Ａ，用 Ｎｈｅ Ｉ、 Ｘｈｏ Ｉ双酶切 Ｐ Ｍ Ａ Ｍ ｎｅｏ５３。采用低熔点琼脂糖凝胶挖块回收法回收 Ｐ１６、 Ｐ５３基因片段，片段长度分别为８００ ｂｐ、１ ８００ ｂｐ。分别经抽提、沉淀、漂洗，使基因片段进一步纯化。利用定向克隆技术将 Ｐ１６、 Ｐ５３基因片段顺向插入经 Ｅｃｏ Ｒ Ｉ、 Ｘｈｏ Ｉ双酶切后的ｐｃ Ｄ Ｎ Ａ３，再次转化后经快速鉴定筛选出 ｐｃ Ｄ Ｎ Ａ１６５３融合表达载体。结果： Ｐ１６、 Ｐ５３的融合表达载体大小８ ０００ ｂｐ、插入方向为正向。结论：用基因重组技术可以将两个抑癌基因构建在一个表达载体中进行表达，这为利用多基因治疗肿瘤的基础研究与临床研究提供了前提条件。
OBJECTIVE: To construct a fusion expression vector containing tumor suppressor gene P16 and P53 by gene recombination. METHODS: P16A was digested with restriction enzyme EcoR I and Xho I, and P M A M neo53 was digested with Nhe I and Xho I. The low-melting point agarose gel excavation recovery method was used to recover the P16 and P53 gene fragments. The fragment lengths were 800 bp and 1 800 bp, respectively. After extraction, precipitation and rinsing, the gene fragments were further purified. Using directional cloning technology, P16 and P53 gene segments were inserted into pcD N A3 digested by EcoR I and Xho I enzymes, and then transformed into PC d N A16-53 fusion expression vector after rapid identification. Results: The size of the fusion expression vector of P16 and P53 was 8 000 bp and the insertion direction was positive. Conclusion: The gene recombination technology can be used to construct two tumor suppressor genes in an expression vector. This provides a prerequisite for the basic research and clinical research of multi-gene therapy for tumors.