M-CSF胞质内稳定表达细胞系的构建和鉴定-CSCIED科技核心评价数据库-手机版

M-CSF胞质内稳定表达细胞系的构建和鉴定

Construction and Identification of M-CSF Stably Expressing Cell Line in Cytoplasm

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DOI	/
刊名	南华大学学报(医学版)
年，卷(期)	2009, 37(02)
作者	涂剑1吴海燕1,2姚志红1朱炳阳1唐圣松1,
作者单位	1.南华大学药物药理研究所2.郴州市第一人民医院病理科
摘要	论文构建真核细胞pCMV/cyto/myc-M-CSF载体,建立M-CSF胞质内稳定表达细胞系,为进一步研究M-CSF的胞内作用奠定基础。结果表明，重组质粒pCMV/cyto/myc-M-CSF用M-CSF特异性的引物进行PCR扩增,结果显示插入片段约为1 400 bp,双酶切后分别得到约5.0 kb和1 400 bp的两条带,M-CSF分子大小与预期一致。RT-PCR、免疫细胞化学结果表明转染M-CSF的HeLa细胞高表达M-CSF(P<0.05),并且转染M-CSF的HeLa细胞表达的M-CSF蛋白定位于细胞质。
Abstract	The study constructed a eukaryotic pCMV/cyto/myc-M-CSF vector and established a stable cell line expressing M-CSF in the cytoplasm, laying a foundation for further investigation into the intracellular functions of M-CSF. The results showed that the recombinant plasmid pCMV/cyto/myc-M-CSF was amplified by PCR using M-CSF-specific primers, yielding an insert fragment of approximately 1,400 bp. Following double enzyme digestion, two bands of about 5.0 kb and 1,400 bp were obtained, consistent with the expected molecular size of M-CSF. RT-PCR and immunocytochemistry results demonstrated that HeLa cells transfected with M-CSF exhibited significantly high expression of M-CSF (P < 0.05), and the expressed M-CSF protein was localized to the cytoplasm.
关键词	巨噬细胞集落刺激因子;靶向定位载体;HeLa细胞;胞质;
KeyWord	macrophage colony-stimulating factor;pCMV/cyto/myc-M-CSF recombinant plasmid;HeLa cells;cytoplasm
基金项目
页码	133-135

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引用本文

涂剑吴海燕,姚志红朱炳阳唐圣松. M-CSF胞质内稳定表达细胞系的构建和鉴定 [J]. 南华大学学报(医学版). 2009; 37; (02). 133 - 135.

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