一种通过热处理提高质粒DNA生物活性的方法

Sen Hou, Kun Yang, Ze Liu, Xi-Zeng Feng
{"title":"一种通过热处理提高质粒DNA生物活性的方法","authors":"Sen Hou,&nbsp;Kun Yang,&nbsp;Ze Liu,&nbsp;Xi-Zeng Feng","doi":"10.1016/j.jprot.2008.01.013","DOIUrl":null,"url":null,"abstract":"<div><p>A method to increase the bioactivity of plasmid DNA by heat treatment has been developed. The structure of the heat treated plasmid DNA was investigated by electrophoresis assay and atomic force microscope (AFM) observation. Electrophoresis assay showed that the heat treated DNA consisted of three components: the supercoiled DNA (component I), the open circular DNA (component II) and the heat denatured DNA component. The bioactivity of the heat treated plasmid DNA was investigated by both DNA condensation experiments and gene transfection experiment with mammal cells. DNA condensation experiments showed that the heat denatured DNA component owned higher sensitivity to spermidine and polyethylenimine (PEI) than component I and component II DNA. Gene transfection experiment with PEI indicated that the heat treated DNA had higher gene transfection efficiency than untreated DNA. Our experiment not only shows an effective approach to increase the bioactivity of plasmid DNA but also leads a way to improve the bioactivity of DNA by physically modifying their structure.</p></div>","PeriodicalId":15257,"journal":{"name":"Journal of biochemical and biophysical methods","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2008-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.jprot.2008.01.013","citationCount":"4","resultStr":"{\"title\":\"A method to increase the bioactivity of plasmid DNA by heat treatment\",\"authors\":\"Sen Hou,&nbsp;Kun Yang,&nbsp;Ze Liu,&nbsp;Xi-Zeng Feng\",\"doi\":\"10.1016/j.jprot.2008.01.013\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>A method to increase the bioactivity of plasmid DNA by heat treatment has been developed. The structure of the heat treated plasmid DNA was investigated by electrophoresis assay and atomic force microscope (AFM) observation. Electrophoresis assay showed that the heat treated DNA consisted of three components: the supercoiled DNA (component I), the open circular DNA (component II) and the heat denatured DNA component. The bioactivity of the heat treated plasmid DNA was investigated by both DNA condensation experiments and gene transfection experiment with mammal cells. DNA condensation experiments showed that the heat denatured DNA component owned higher sensitivity to spermidine and polyethylenimine (PEI) than component I and component II DNA. Gene transfection experiment with PEI indicated that the heat treated DNA had higher gene transfection efficiency than untreated DNA. Our experiment not only shows an effective approach to increase the bioactivity of plasmid DNA but also leads a way to improve the bioactivity of DNA by physically modifying their structure.</p></div>\",\"PeriodicalId\":15257,\"journal\":{\"name\":\"Journal of biochemical and biophysical methods\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2008-04-24\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1016/j.jprot.2008.01.013\",\"citationCount\":\"4\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of biochemical and biophysical methods\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0165022X07002059\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of biochemical and biophysical methods","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0165022X07002059","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 4

摘要

提出了一种通过热处理提高质粒DNA生物活性的方法。用电泳法和原子力显微镜观察热处理后质粒DNA的结构。电泳分析表明,热处理后的DNA由超螺旋DNA(组分I)、开环DNA(组分II)和热变性DNA(组分II)三部分组成。通过DNA浓缩实验和哺乳动物细胞基因转染实验,研究了热处理质粒DNA的生物活性。DNA缩合实验表明,热变性DNA组分对亚精胺和聚乙烯亚胺(PEI)的敏感性高于组分I和组分II DNA。PEI基因转染实验表明,热处理DNA的基因转染效率高于未处理DNA。本实验不仅为提高质粒DNA的生物活性提供了一条有效途径,而且为通过物理修饰DNA的结构来提高DNA的生物活性开辟了一条途径。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
A method to increase the bioactivity of plasmid DNA by heat treatment

A method to increase the bioactivity of plasmid DNA by heat treatment has been developed. The structure of the heat treated plasmid DNA was investigated by electrophoresis assay and atomic force microscope (AFM) observation. Electrophoresis assay showed that the heat treated DNA consisted of three components: the supercoiled DNA (component I), the open circular DNA (component II) and the heat denatured DNA component. The bioactivity of the heat treated plasmid DNA was investigated by both DNA condensation experiments and gene transfection experiment with mammal cells. DNA condensation experiments showed that the heat denatured DNA component owned higher sensitivity to spermidine and polyethylenimine (PEI) than component I and component II DNA. Gene transfection experiment with PEI indicated that the heat treated DNA had higher gene transfection efficiency than untreated DNA. Our experiment not only shows an effective approach to increase the bioactivity of plasmid DNA but also leads a way to improve the bioactivity of DNA by physically modifying their structure.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
自引率
0.00%
发文量
0
期刊最新文献
Editorial Board Fluorescent method for detection of cleaved collagens using O-phthaldialdehyde (OPA) A rapid and non leaky way for preparation of the sharp intracellular recording microelectrodes Quantification of penicillin G during labor and delivery by capillary electrophoresis Dispensing an enzyme-conjugated solution into an ELISA plate by adapting ink-jet printers
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1