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RT PCR扩增获得mSemcap2全长基因并在真核细胞中表达 ,对其功能进行初步研究。用DNA重组法构建了真核表达质粒pEGFP N1 Semcap2。将重组质粒利用电穿孔法转入HeLa细胞中 ,通过G4 18进行压力筛选。稳定表达mSemcap2的细胞株与PP结 (Peyer’spatch )淋巴细胞共培养 2d后通过FACS观察其对淋巴细胞表型的影响。结果是mSemcap2在真核细胞中得到了成功的表达。稳定表达mSemcap2基因的细胞株可改变与之共培养的PP结淋巴细胞的表型 ,使B2 2 0 + 细胞增多。说明mSemcap2分子可能与免疫系统功能相关 ;其稳定表达细胞株的获得为深入研究mSemcap2的功能提供了材料。
The full-length mSemcap2 gene was amplified by RT-PCR and expressed in eukaryotic cells. The function of mSemcap2 gene was preliminarily studied. The eukaryotic expression plasmid pEGFP N1 Semcap2 was constructed by DNA recombination. The recombinant plasmids were electroporated into HeLa cells and pressure-screened by G418. Cell lines stably expressing mSemcap2 were co-cultured with Peyer’s patch lymphocytes for 2 days and their effects on lymphocyte phenotypes were observed by FACS. The result is that mSemcap2 has been successfully expressed in eukaryotic cells. Cell lines stably expressing the mSemcap2 gene can alter the phenotype of PP-conjuged lymphocytes co-cultured with the B220 + cells. These results suggest that mSemcap2 may be involved in the function of immune system. The expression of mSemcap2 in stable cell lines may provide some useful information for further study on the function of mSemcap2.