HSF1基因敲除小鼠胚胎成纤维细胞的永生化
目的:建立HSF1^-/-,HSF1^+/+两种基因型小鼠胚胎成纤维永生化细胞系,为HSF1的功能研究提供实验模型。方法:用脂质体介导的基因转染法将pSV3neo质粒导入HSF1^-/-,HSF1^+/+两种基因型小鼠胚胎成纤维细胞,经G418筛选,抗性克隆扩大培养,建立永生化细胞系;用PCR检测两种细胞株中目的基因的整合,用RT—PCR法鉴定SV40T基因在转染细胞中的表达;用Western blot检测所建细胞株的诱导型热休克蛋白70的表达情况。结果:有3个细胞克隆已扩大培养稳定传代达6个月,经鉴定SV40T抗原已整合到两种细胞中且稳定表达,HSF1^-/-胚胎成纤维细胞热休克蛋白70的诱...
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Veröffentlicht in: | Zhong nan da xue xue bao. Journal of Central South University. Yi xue ban 2006, Vol.31 (2), p.174-177 |
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container_title | Zhong nan da xue xue bao. Journal of Central South University. Yi xue ban |
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creator | 刘梅冬 张华莉 龚环宇 陈广文 王慷慨 鄂顺梅 肖献忠 |
description | 目的:建立HSF1^-/-,HSF1^+/+两种基因型小鼠胚胎成纤维永生化细胞系,为HSF1的功能研究提供实验模型。方法:用脂质体介导的基因转染法将pSV3neo质粒导入HSF1^-/-,HSF1^+/+两种基因型小鼠胚胎成纤维细胞,经G418筛选,抗性克隆扩大培养,建立永生化细胞系;用PCR检测两种细胞株中目的基因的整合,用RT—PCR法鉴定SV40T基因在转染细胞中的表达;用Western blot检测所建细胞株的诱导型热休克蛋白70的表达情况。结果:有3个细胞克隆已扩大培养稳定传代达6个月,经鉴定SV40T抗原已整合到两种细胞中且稳定表达,HSF1^-/-胚胎成纤维细胞热休克蛋白70的诱导表达消失。结论:成功建立永生化HSF1^-/-,HSF1^+/+两种基因型小鼠胚胎成纤维细胞。 |
doi_str_mv | 10.3321/j.issn:1672-7347.2006.02.004 |
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Yi xue ban</title><addtitle>Journal of Central South University (Medical Sciences) Journal of Central South University (Medical Sciences)</addtitle><description>目的:建立HSF1^-/-,HSF1^+/+两种基因型小鼠胚胎成纤维永生化细胞系,为HSF1的功能研究提供实验模型。方法:用脂质体介导的基因转染法将pSV3neo质粒导入HSF1^-/-,HSF1^+/+两种基因型小鼠胚胎成纤维细胞,经G418筛选,抗性克隆扩大培养,建立永生化细胞系;用PCR检测两种细胞株中目的基因的整合,用RT—PCR法鉴定SV40T基因在转染细胞中的表达;用Western blot检测所建细胞株的诱导型热休克蛋白70的表达情况。结果:有3个细胞克隆已扩大培养稳定传代达6个月,经鉴定SV40T抗原已整合到两种细胞中且稳定表达,HSF1^-/-胚胎成纤维细胞热休克蛋白70的诱导表达消失。结论:成功建立永生化HSF1^-/-,HSF1^+/+两种基因型小鼠胚胎成纤维细胞。</description><subject>HSF1</subject><subject>SV40</subject><subject>T抗原</subject><subject>基因敲除</subject><subject>永生化</subject><subject>胚胎成纤维细胞</subject><issn>1672-7347</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2006</creationdate><recordtype>article</recordtype><recordid>eNo9j81Kw0AUhWehYKl9CRFcJU7m3mQy7qRYKxRc2H1IZpI2bUnRIOo6Ii5EXahgBdtFH6CIC2l9nfz0LYxU3JwDh49z7yFk26A6ADN2e3oYx9GeYXGmcUCuM0otnTKdUlwjlf98g9TiOPQoFUKAZfMK4c2ThpGO5-nbJHv-WL5O09nD8ntSJKMiuc9uH_P5NF985oubInnPR9fZ7Ct_Gqd3L5tkPXAHsV_78yppNw7a9abWOj48qu-3NGkK1LhiykPlmZaFoJR00QwkN22OHvMBbaSSIYBpK8m5GTAmlI0281FIBlD-WCU7q9oLNwrcqOP0hudnUXnQ6UZXfXX5u5OWgiW5tSJldxh1TsOS9VzZD8KB7zCDg-AC4Acks2bM</recordid><startdate>2006</startdate><enddate>2006</enddate><creator>刘梅冬 张华莉 龚环宇 陈广文 王慷慨 鄂顺梅 肖献忠</creator><general>中南大学,湘雅医学院病理生理学教研室,长沙,410078%中南大学,湘雅三医院,长沙,410013</general><scope>2RA</scope><scope>92L</scope><scope>CQIGP</scope><scope>W94</scope><scope>WU4</scope><scope>~WA</scope><scope>2B.</scope><scope>4A8</scope><scope>92I</scope><scope>93N</scope><scope>PSX</scope><scope>TCJ</scope></search><sort><creationdate>2006</creationdate><title>HSF1基因敲除小鼠胚胎成纤维细胞的永生化</title><author>刘梅冬 张华莉 龚环宇 陈广文 王慷慨 鄂顺梅 肖献忠</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c594-7d2db4db56643ddca45fc75874b2e34840c243358dc775f229d8482e49c233993</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>chi</language><creationdate>2006</creationdate><topic>HSF1</topic><topic>SV40</topic><topic>T抗原</topic><topic>基因敲除</topic><topic>永生化</topic><topic>胚胎成纤维细胞</topic><toplevel>online_resources</toplevel><creatorcontrib>刘梅冬 张华莉 龚环宇 陈广文 王慷慨 鄂顺梅 肖献忠</creatorcontrib><collection>中文科技期刊数据库</collection><collection>中文科技期刊数据库-CALIS站点</collection><collection>中文科技期刊数据库-7.0平台</collection><collection>中文科技期刊数据库-自然科学</collection><collection>中文科技期刊数据库-自然科学-生物科学</collection><collection>中文科技期刊数据库- 镜像站点</collection><collection>Wanfang Data Journals - Hong Kong</collection><collection>WANFANG Data Centre</collection><collection>Wanfang Data Journals</collection><collection>万方数据期刊 - 香港版</collection><collection>China Online Journals (COJ)</collection><collection>China Online Journals (COJ)</collection><jtitle>Zhong nan da xue xue bao. 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Yi xue ban</jtitle><addtitle>Journal of Central South University (Medical Sciences) Journal of Central South University (Medical Sciences)</addtitle><date>2006</date><risdate>2006</risdate><volume>31</volume><issue>2</issue><spage>174</spage><epage>177</epage><pages>174-177</pages><issn>1672-7347</issn><abstract>目的:建立HSF1^-/-,HSF1^+/+两种基因型小鼠胚胎成纤维永生化细胞系,为HSF1的功能研究提供实验模型。方法:用脂质体介导的基因转染法将pSV3neo质粒导入HSF1^-/-,HSF1^+/+两种基因型小鼠胚胎成纤维细胞,经G418筛选,抗性克隆扩大培养,建立永生化细胞系;用PCR检测两种细胞株中目的基因的整合,用RT—PCR法鉴定SV40T基因在转染细胞中的表达;用Western blot检测所建细胞株的诱导型热休克蛋白70的表达情况。结果:有3个细胞克隆已扩大培养稳定传代达6个月,经鉴定SV40T抗原已整合到两种细胞中且稳定表达,HSF1^-/-胚胎成纤维细胞热休克蛋白70的诱导表达消失。结论:成功建立永生化HSF1^-/-,HSF1^+/+两种基因型小鼠胚胎成纤维细胞。</abstract><pub>中南大学,湘雅医学院病理生理学教研室,长沙,410078%中南大学,湘雅三医院,长沙,410013</pub><doi>10.3321/j.issn:1672-7347.2006.02.004</doi><tpages>4</tpages></addata></record> |
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subjects | HSF1 SV40 T抗原 基因敲除 永生化 胚胎成纤维细胞 |
title | HSF1基因敲除小鼠胚胎成纤维细胞的永生化 |
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