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pcDNA3.1一MORC2重组质粒的构建及其在胃癌细胞SGC-7901中的表达和定位
  • ISSN号:1006-2947
  • 期刊名称:《解剖科学进展》
  • 时间:0
  • 分类:R735.2[医药卫生—肿瘤;医药卫生—临床医学]
  • 作者机构:[1]中国医科大学,细胞生物学教研室,卫生部细胞生物学重点实验室,沈阳110001
  • 相关基金:国家自然科学基金资助项目(No.30370736,No.30570966)、教育部博士点基金资助项目(No.20050159023)
中文摘要:

目的构建真核表达载体pcDNA3.1-MORC2并瞬时转染人胃癌细胞SGC-7901,观察融合蛋白在细胞内表达及定位。方法以人MORC2cDNA为模板,PCR扩增MORC2全长编码基因,亚克隆至pcDNA3.1-hisA表达载体。将构建的重组质粒测序并转染到胃癌细胞SGC-7901中,提取细胞蛋白进行Westernblot检测。用共聚焦激光扫描显微镜观察pcDNA3.1-MORC2在SGC-7901细胞内定位。结果MORC2全长基因序列克隆到了表达载体pcDNA3.1-hisAr中,酶切鉴定片段为3000bp。Westernblot检测到了融合蛋白pcDNA3.1-MORC2在胃癌细胞系SGC-7901中表达,分子量约为122kDa,主要定位于细胞核。结论成功构建了真核表达载体pcDNA3.1-MORC2,并检测到融合蛋白表达主要定位于胃癌细胞核内。

英文摘要:

Objective To construct the recombinant expression plasmid of microchidia2 (MORC2) gene and identify its protein expression and localization in SGC-7901 cells. Methods The MORC2 coding sequence was amplified by polymerase chain reaction (PCR) method and subcloned into pcDNA3.1-hisA vector. After the target region was sequenced, the plasmid was transfected into SGC-7901 cell lines. The expression of the recombinant plasmid in SGC-7901 cells was proved by Western blot and the localization of pcDNA3.1-morc2 was observed by using laser scanning confocal microscopy. Results MORC2 was constructed into expressing vector pcDNA3.1-hisA successfully, the length of the fragment was 3000bp, identified by restriction enzymes digestion. The expression of pcDNA3.1-MORC2 fusion protein was detected by Western blot in SGC-7901 cells with a molecular weight 122kDa, and localized in the nucleus of SGC-7901 cells. Conclusions The recombinant plasmid of pcDNA3.1-MORC2 was constructed successfully and the fusion protein was localized in nucleus of SGC-7901 ceils.

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期刊信息
  • 《解剖科学进展》
  • 中国科技核心期刊
  • 主管单位:中国科学技术协会
  • 主办单位:中国解剖学会
  • 主编:方秀斌
  • 地址:沈阳市和平北二马路92号中国医科大学内
  • 邮编:110001
  • 邮箱:
  • 电话:024-31939627
  • 国际标准刊号:ISSN:1006-2947
  • 国内统一刊号:ISSN:21-1347/Q
  • 邮发代号:8-116
  • 获奖情况:
  • 国内外数据库收录:
  • 被引量:6163