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目的 构建可应用于昆虫杆状病毒表达系统的含抑癌基因p16 INK4cDNA的重组转移载体。方法 采用定向克隆方法将抑癌基因p16 INK4cDNA全长插入转移载体质粒pSXIVVI+ X3,并经斑点杂交 ,PCR及酶切鉴定重组质粒。结果 经三种鉴定方法证实p16 INK4cDNA片段成功地插入转移载体pSXIVVI+ X3。结论 重组质粒pSXIVVI+ X3 p16构建成功 ,为真核表达抑癌基因p16 INK4奠定了基础。
Objective To construct a recombinant transfer vector containing tumor suppressor gene p16 INK4 cDNA that can be used in insect baculovirus expression system. METHODS: The full-length anti-oncogene p16 INK4 cDNA was inserted into the transfer vector plasmid pSXIVVI+X3 by directional cloning, and the recombinant plasmid was identified by dot blot, PCR and restriction endonuclease digestion. Results Three identification methods confirmed that the p16 INK4 cDNA fragment was successfully inserted into the transfer vector pSXIVVI+X3. Conclusion The successful construction of the recombinant plasmid pSXIVVI+X3 p16 lays the foundation for the eukaryotic expression of tumor suppressor gene p16 INK4.