M-CSF核内稳定表达细胞系的构建和鉴定
Construction of nuclear localization vector pCMV/M-CSF and establishment of its stably expressing cell line
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| DOI |
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| 刊名 |
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| 年，卷(期) |
2009, 25(03) |
| 作者 |
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| 作者单位 |
1.南华大学药物药理研究所2.岳阳职业技术学院3.南华大学组织胚胎学教研室
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| 摘要 |
论文构建真核细胞pCMV/nuc/M-CSF载体,建立M-CSF核内稳定表达细胞系,为进一步研究M-CSF的核内作用奠定基础。结果表明，琼脂糖电泳结果显示插入片段为1400bp左右,与预期M-CSF分子大小相当;DNA测序分析表明插入质粒的M-CSF无读码框移位,并与来源序列一致。RT-PCR和间接免疫荧光检测表明,转染pCMV/M-CSF的HeLa细胞能稳定表达M-CSFmRNA与M-CSF蛋白,且表达的M-CSF定位于HeLa细胞的细胞核。
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| Abstract |
The study constructed a eukaryotic cell pCMV/nuc/M-CSF vector and established an M-CSF nuclear stable expression cell line, laying the foundation for further research on the nuclear function of M-CSF. The results showed that agarose gel electrophoresis indicated an inserted fragment of approximately 1400 bp, which corresponded to the expected molecular size of M-CSF. DNA sequencing analysis confirmed that the M-CSF inserted into the plasmid had no frameshift mutations and was consistent with the original sequence. RT-PCR and indirect immunofluorescence assays demonstrated that HeLa cells transfected with pCMV/M-CSF stably expressed M-CSF mRNA and M-CSF protein, with the expressed M-CSF localized to the nucleus of the HeLa cells.
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| 关键词 |
巨噬细胞集落刺激因子;pCMV/myc/nuc载体;HeLa细胞;
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| KeyWord |
Macrophage colony-stimulating factor; pCMV/myc/nuc vector; HeLa cells
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| 基金项目 |
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| 页码 |
622-624 |
涂剑滕淑静张晓红赵雪琴唐圣松.
M-CSF核内稳定表达细胞系的构建和鉴定 [J].
中国病理生理杂志.
2009; 25; (03).
622 - 624.