CAJ | 학술논문

目的：利用乳链菌穿梭载体构建肿瘤特异标记表达系统，为肿瘤特异性口服疫苗的研究开辟一个新领域。方法采用体外基因拼装的方法得到PEG-3启动子，经酶切鉴定并测序验证。然后利用基因克隆技术，将PEG-3启动子、EGFP、Poly A亚克隆到乳链菌载体pTRKH2中，成功得到肿瘤特异标记质粒pTR-peg3-egfp。最后，转染肿瘤及非肿瘤细胞初步验证其肿瘤特异性标记能力。结果通过PCR基因拼装得到460bp左右目的片段，测序结果与设计完全一致。获得了肿瘤特异标记的乳链菌穿梭质粒pTR-peg3-egfp，并在细胞水平得到初步验证。结论 PEG-3启动子介导的乳链菌穿梭质粒具有肿瘤特异性，这一实用新颖的系统将为研究乳链菌疫苗应用于肿瘤标记及靶向治疗奠定基础。
목적：이용유련균천사재체구건종류특이표기표체계통，위종류특이성구복역묘적연구개벽일개신영역。방법채용체외기인병장적방법득도PEG-3계동자，경매절감정병측서험증。연후이용기인극륭기술，장PEG-3계동자、EGFP、Poly A아극륭도유련균재체pTRKH2중，성공득도종류특이표기질립pTR-peg3-egfp。최후，전염종류급비종류세포초보험증기종류특이성표기능력。결과통과PCR기인병장득도460bp좌우목적편단，측서결과여설계완전일치。획득료종류특이표기적유련균천사질립pTR-peg3-egfp，병재세포수평득도초보험증。결론 PEG-3계동자개도적유련균천사질립구유종류특이성，저일실용신영적계통장위연구유련균역묘응용우종류표기급파향치료전정기출。
Objective The aim of this study is to construct a tumor-specific imaging system with the shuttle vector of lactococcus lacti which will open a new field for the study of tumor-specific oral vaccine. Methods The PEG-3 promoter was constructed by means of PCR base gene assembly, and conformed by en-zyme digestion and DNA sequencing. Then, constructed recombinant pTR-peg3-egfp (shuttle plasmid) carry-ing the PEG-3 promoter enhanced green fluorescent protein and Poly A tail. Finally, the ability of tumor-spe-cific imaging in the means of in vitro transfection was validated. Results A 460 bp purpose fragment obtained by PCR gene splicing agreed exactly with the design. The tumor specific lactococcus lacti plasmid pTR-peg3-egfp was successfully constructed , and also got a preliminary validation on cellular level. Conclusion Lacto-coccus lacti vector with PEG-3 promoter could specifically recognize tumor cell. This utility, new lactococ-cus lacti system can lay a solid foundation for both cancer imaging and tumor-specific gene therapy.