摘要
目的克隆出针对表皮生长因子受体Ⅲ型突变体(EGFRvⅢ)PEP-3表位的cDNA片段,为高表达EGFRvⅢ肿瘤的免疫治疗奠定基础。方法根据PEP-3的碱基序列设计出有12对互补碱基的正负引物,正负引物互为模板进行PCR扩增,制备出两端含酶切位点的PEP-3 cDNA片段,再将扩增的PCR产物亚克隆于载体pGEM-T Easy构建重组质粒pGEM-T Easy/PEP-3。结果经限制性内切酶酶切鉴定和DNA测序证实,编码PEP-3的cDNA片段被成功扩增并亚克隆于载体pGEM-T Easy中。结论成功克隆了编码表皮生长因子受体Ⅲ型突变体PEP-3表位的cDNA片段。
Objective Through cloning the cDNA fragment encoding PEP-3 epitope of epidermal growth factor receptor type Ⅲ mutant（EGFRvⅢ） to establish the base for immunotherapy of carcinoma with high EGFRv Ⅲ expressing. Methods Designing the primer according to the base sequence of PEP-3, cDNA was synthesized and amplificated by PCR. Then, the PCR product was subcloned into vector pGEM-T Easy so as to construct recombinant plasmid pGEM-T Easy/ PEP-3. Results Zymo-cutting restriction enzyme identification and DNA sequencing conformed that cDNA encoding PEP-3 was amplified and subcoloned into pGEM-T Easy successfully. Conclusion cDNA encoding PEP-3 epitope of EGFRvⅢ was coloned successfully.
出处
《西安交通大学学报（医学版）》
CAS
CSCD
北大核心
2006年第2期140-142,共3页
Journal of Xi’an Jiaotong University（Medical Sciences）
