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Isolation and cloning of sense and antisense K-ras gene fragments
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- Author:
- No author available
- Journal Title:
- CHINESE JOURNAL OF PANCREATOLOGY
- Issue:
- 4
- DOI:
- 10.3760/cma.j.issn.1674-1935.2003.04.004
- Key Word:
- 胰腺肿瘤;基因,ras;癌基因遗传载体;逆转录病毒科;转导,遗传
Abstract： 目的分离及克隆胰腺癌基因组中K-ras基因片段,构建重组正反义K-ras基因逆转录病毒载体并探讨其临床意义.方法设计两对PCR引物,分别在上下游引物中引进BamH1和EcoR1位点,以胰腺癌细胞基因组DNA为模板扩增K-ras基因外显子4B及侧翼序列,并采用重组DNA技术将目的基因分别定向插入逆转录病毒载体LZRSpBMN-Z中.重组克隆通过菌液PCR和重组质粒限制性内切酶酶切鉴定.结果正反义K-ras基因外显子4B及侧翼序列成功地克隆入LZRSpBMN-Z中.结论LZRSpBMN-Z是胰腺癌反义基因治疗中新型候选载体之一.应用PCR方法获取反义K-ras基因方便可行,可用于重组反义K-ras基因逆转录病毒载体的构建.
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