CAJ | 학술논문

目的：检测真核质粒pMAGEA3-IRES-SEA经过电穿孔转染在小鼠中的转录。方法将葡萄球菌肠毒素A（staphylococcal endotoxin A，SEA）和喉癌来源的黑色素瘤抗原A3（melanoma-associated antigen A3， MAGE-A3），构建成基因共表达的真核质粒 pMAGEA3-IRES-SEA。将pMAGEA3-IRES-SEA用电转染的方法免疫BALB/C小鼠单侧股四头肌，每2周1次，每次注射50μg，共免疫3次。末次免疫2周后，取注射部位组织，用荧光定量PCR（qRT-PCR）法检测目的基因在注射部位肌肉中的转录情况。结果在实验小鼠注射部位的骨骼肌内检测到 SEA、MAGE-A3基因转录mRNA。结论所构建的pMAGEA3-IRES-SEA真核表达质粒，可通过电转染的方式，在小鼠体内能有效地转录。这为研究该质粒通过小鼠的体内转录蛋白的表达，诱导机体细胞免疫、体液免疫来清除喉癌，奠定了理论基础。
목적：검측진핵질립pMAGEA3-IRES-SEA경과전천공전염재소서중적전록。방법장포도구균장독소A（staphylococcal endotoxin A，SEA）화후암래원적흑색소류항원A3（melanoma-associated antigen A3， MAGE-A3），구건성기인공표체적진핵질립 pMAGEA3-IRES-SEA。장pMAGEA3-IRES-SEA용전전염적방법면역BALB/C소서단측고사두기，매2주1차，매차주사50μg，공면역3차。말차면역2주후，취주사부위조직，용형광정량PCR（qRT-PCR）법검측목적기인재주사부위기육중적전록정황。결과재실험소서주사부위적골격기내검측도 SEA、MAGE-A3기인전록mRNA。결론소구건적pMAGEA3-IRES-SEA진핵표체질립，가통과전전염적방식，재소서체내능유효지전록。저위연구해질립통과소서적체내전록단백적표체，유도궤체세포면역、체액면역래청제후암，전정료이론기출。
Objective To identify the transcription of eukaryotic coexpression vector pMAGEA3-IRES-SEA by electroporation method in muscle of mice. Methods The eukaryotic coexpression vector was constructed with staphylococcal endotoxin A(SEA) and human melanoma-associated antigen gene A3 (MAGE-A3) originating from laryngocarcinoma by genetic recombinant method in advance. Unilateral hindlimb skeletal muscles of BALB/C mice were immuned with eukaryotic expression plasmid pMAGEA3-IRES-SEA by electrotransfection method,one time every two week, each immuned of 50μg, a total of three times. Two weeks after the final immunization, their gene transcriptions were identified at the muscle organization of the injection site by fluorescence quantitative PCR (qRT-PCR) assay. Results The SEA and MAGE-A3 genes had been transcripted in experimental mice skeletal muscle. Conclusions The constructed pMAGEA3-IRES-SEA eukaryotic expression plasmid can be effectively transcripted in mice by electroporation method. This work provided a theoretical basis for the expression of mouse in vivo transcription protein and the application of the plasmid to clear laryngocarcinoma by inducing the cellular immunity and humoral immunity in organism.