| 特异性DNA倍增技术检测t-PA cDNA基因在表达细胞中的稳定性 |
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| 引用本文： | 付四清,严建,龚小华,翁致平,宋后燕. 特异性DNA倍增技术检测t-PA cDNA基因在表达细胞中的稳定性[J]. 复旦学报(医学版), 1990, 0(5) |
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| 作者姓名： | 付四清 严建 龚小华 翁致平 宋后燕 |
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| 作者单位： | 上海医科大学病理生理学教研室(付四清)，中山医院心血管病研究所(严建)，上海医科大学基础医学部分子遗传学(龚小华，翁致平)，上海医科大学基础医学部分子遗传学(宋后燕) |
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| 基金项目： | 国家“八六三”计划资助课题 |
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| 摘 要： | 利用特异性DNA倍增技术(polymerase chain Reaction,PCR)检测t-PA cDNA基因在表达细胞基因组中的稳定性,并对所得的PCR反应产物进行了限制性内切酶片段、分子杂交和核苷酸顺序分析等方面的研究,证实了t-PA cDNA基因已插入到表达细胞染色体中。这种方法快速、简便、灵敏度和特异性高,是检测基因工程表达细胞中cDNA基因稳定整合状况的好方法。
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| 关 键 词： | 特异性DNA倍增技术 组织纤溶酶原激活剂 cDNA稳定整合 |
DETECTION OF THE STABILITY OF t-PA cDNA IN EXPRESSION CELLS WITH THE POLYMERASE CHAIN REACTION |
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| Fu Siqing,Yan Jian,Gong Xlaohua,Weng Zhiping,Song Houyan. DETECTION OF THE STABILITY OF t-PA cDNA IN EXPRESSION CELLS WITH THE POLYMERASE CHAIN REACTION[J]. Fudan University Journal of Medical Sciences, 1990, 0(5) |
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| Authors: | Fu Siqing Yan Jian Gong Xlaohua Weng Zhiping Song Houyan |
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| Abstract: | The polymerase chain reaction (PCK) technique was applied to ascertain the Integration of tissue plasminogen activator (t-PA) oDNA into the genome in t-PA oDNA expression cells. Gel eleotrophoresis, analysis of restriction endonuolease pragments, hybridization with a t-PA oDNA probe and DNA sequencing of the PCR froduot verified t-PA oDNA gene integration into the cellular genome of t-PA oDNA transfeoted cells and ita stability. The PCR technique is rapid, simple, highly-sensitive and highly-specific for ascertaining the stability of the oDNA in transfected cells. |
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| Keywords: | polymarase chain reaction tissue plasminogen activator stability of eDNA integration |
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