摘要
目的:分离及克隆胰腺癌基因组中K-ras基因片段,构建重组反义K-ras癌基因逆转录病毒载体,并探讨其意义。方法:设计2对PCR引物,分别在上下游引物中引进BamHⅠ和EcoRⅠ位点,以胰腺癌细胞株BxPC-3基因组DNA为模板扩增K-ras基因外显子1及侧翼序列,并采用重组DNA技术将目的基因插入逆转录病毒载体LZRSpBMN-Z中,并经菌液PCR和限制性内切酸酶切鉴定。结果:K-ras基因外显子1及侧翼序列已成功地克隆入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目的基因外显子1方便可行,可用于重组反义K-ras癌基因逆转录病毒载体的构建。
Objective: To isolate and clone antisense K-ras gene fragment in pancreatic carcinoma. Methods: Polymerase chain re-action(PCR) was used to amplify exon 1 and flanking sequence of BxPC-3 pancreatic carcinoma cell line genomic DNA and the PCR product was inserted into retrovial vector LZRSpBMN-Z using recombinant DNA technique. The recombinant plasmid was isolated, purified and identified by PCR and restriction enzyme digestion. Results:The exon 1 and flanking sequence of K-ras gene were successfully cloned into the vector LZRSpBMN-Z. Conclusion: The vector LZRSpBMN-Z may be a novel ideal candidate vector for gene therapy of pancreatic carcinoma. Antisense K-ras gene fragment cloned by PCR could be applied to construct of retroviral vector of recombinant.
出处
《南京医科大学学报（自然科学版）》
CAS
CSCD
北大核心
2003年第6期554-557,共4页
Journal of Nanjing Medical University(Natural Sciences)
