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Gene targeting in murine embryonic stem cells: Introduction of specific alterations into the mammalian genome 小鼠胚胎干细胞的基因靶向:将特异性改变引入哺乳动物基因组
Gene analysis techniques Pub Date : 1990-12-01 DOI: 10.1016/0735-0651(90)90004-Y
Suzanne L Mansour
{"title":"Gene targeting in murine embryonic stem cells: Introduction of specific alterations into the mammalian genome","authors":"Suzanne L Mansour","doi":"10.1016/0735-0651(90)90004-Y","DOIUrl":"10.1016/0735-0651(90)90004-Y","url":null,"abstract":"<div><p>The ability to create targeted mutations in the mouse will have an impact on many areas of research in mammalian biology. Mutations are generated in embryonic stem (ES) cells by homologous recombination between exogenously added DNA and the endogenous chromosomal sequences. These cells are then used to generate chimeric intermediates that pass the mutant allele through the germ line, initiating a strain of mice that carry the desired mutation. This review focuses on the selection of a starting ES cell line, introduction of DNA into ES cells, construction of gene targeting vectors, and selection/enrichment schemes for the isolation of targeted cell lines. The generation of mice that carry the targeted allele is briefly outlined.</p></div>","PeriodicalId":77714,"journal":{"name":"Gene analysis techniques","volume":"7 8","pages":"Pages 219-227"},"PeriodicalIF":0.0,"publicationDate":"1990-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0735-0651(90)90004-Y","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"13247327","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 31
Author index volume 7 作者索引第7卷
Gene analysis techniques Pub Date : 1990-12-01 DOI: 10.1016/0735-0651(90)90006-2
{"title":"Author index volume 7","authors":"","doi":"10.1016/0735-0651(90)90006-2","DOIUrl":"https://doi.org/10.1016/0735-0651(90)90006-2","url":null,"abstract":"","PeriodicalId":77714,"journal":{"name":"Gene analysis techniques","volume":"7 8","pages":"Page 234"},"PeriodicalIF":0.0,"publicationDate":"1990-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0735-0651(90)90006-2","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71784099","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Subject index volume 7 主题索引第7卷
Gene analysis techniques Pub Date : 1990-12-01 DOI: 10.1016/0735-0651(90)90007-3
{"title":"Subject index volume 7","authors":"","doi":"10.1016/0735-0651(90)90007-3","DOIUrl":"https://doi.org/10.1016/0735-0651(90)90007-3","url":null,"abstract":"","PeriodicalId":77714,"journal":{"name":"Gene analysis techniques","volume":"7 8","pages":"Pages 235-237"},"PeriodicalIF":0.0,"publicationDate":"1990-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0735-0651(90)90007-3","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71784100","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
The technology and literature of molecular genetics 分子遗传学的技术和文献
Gene analysis techniques Pub Date : 1990-12-01 DOI: 10.1016/0735-0651(90)90001-V
Glen A Evans (Executive Editor)
{"title":"The technology and literature of molecular genetics","authors":"Glen A Evans (Executive Editor)","doi":"10.1016/0735-0651(90)90001-V","DOIUrl":"https://doi.org/10.1016/0735-0651(90)90001-V","url":null,"abstract":"","PeriodicalId":77714,"journal":{"name":"Gene analysis techniques","volume":"7 8","pages":"Pages 209-210"},"PeriodicalIF":0.0,"publicationDate":"1990-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0735-0651(90)90001-V","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71784493","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
GATA referees 1989–1990
Gene analysis techniques Pub Date : 1990-12-01 DOI: 10.1016/0735-0651(90)90002-W
{"title":"GATA referees 1989–1990","authors":"","doi":"10.1016/0735-0651(90)90002-W","DOIUrl":"https://doi.org/10.1016/0735-0651(90)90002-W","url":null,"abstract":"","PeriodicalId":77714,"journal":{"name":"Gene analysis techniques","volume":"7 8","pages":"Page 211"},"PeriodicalIF":0.0,"publicationDate":"1990-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0735-0651(90)90002-W","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71784494","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
A solution hybridization method for quantification of mRNAs: Determining the amount and stability of oncogene mRNA 一种定量mRNA的溶液杂交法:测定致癌基因mRNA的数量和稳定性
Gene analysis techniques Pub Date : 1990-12-01 DOI: 10.1016/0735-0651(90)90005-Z
Jukka Tenhunen , Jyrki Eloranta, Arja Kallio, Hans Söderlund
{"title":"A solution hybridization method for quantification of mRNAs: Determining the amount and stability of oncogene mRNA","authors":"Jukka Tenhunen ,&nbsp;Jyrki Eloranta,&nbsp;Arja Kallio,&nbsp;Hans Söderlund","doi":"10.1016/0735-0651(90)90005-Z","DOIUrl":"10.1016/0735-0651(90)90005-Z","url":null,"abstract":"<div><p>A solution hybridization method for the quantification of specific mRNAs is described. This assay utilizes complementary RNA probes prepared by in vitro transcription, sandwich hybridization in solution, and affinity-based hybrid collection. The possibility of using this method for crude biological samples without purifying mRNAs makes it ideal when accurate quantification of multiple samples is needed. Human <em>N</em>-myc oncogene transcript was used as a model and as little as 0.24 pg (2 × 10<sup>5</sup> molecules) of N-<em>myc</em> mRNA could be detected. Using this assay it was shown that human neuroblastoma IMR-32 cells contain ′500 N-<em>myc</em> mRNA molecules per cell having a half-life of ′35 min.</p></div>","PeriodicalId":77714,"journal":{"name":"Gene analysis techniques","volume":"7 8","pages":"Pages 228-233"},"PeriodicalIF":0.0,"publicationDate":"1990-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0735-0651(90)90005-Z","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"13247328","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 10
The use of transgenic mice for short-term, in vivo mutagenicity testing 利用转基因小鼠进行短期体内诱变试验
Gene analysis techniques Pub Date : 1990-12-01 DOI: 10.1016/0735-0651(90)90003-X
Steven W. Kohler , G.Scott Provost , Patricia L. Kretz , Annabeth Fieck , Joseph A. Sorge , Jay M. Short
{"title":"The use of transgenic mice for short-term, in vivo mutagenicity testing","authors":"Steven W. Kohler ,&nbsp;G.Scott Provost ,&nbsp;Patricia L. Kretz ,&nbsp;Annabeth Fieck ,&nbsp;Joseph A. Sorge ,&nbsp;Jay M. Short","doi":"10.1016/0735-0651(90)90003-X","DOIUrl":"10.1016/0735-0651(90)90003-X","url":null,"abstract":"<div><p>In order to develop a short-term, in vivo assay to study the mutagenic effects of chemical exposure, transgenic mice were generated using a lambda shuttle vector containing a <em>lac</em>Z target gene. Following exposure to mutagens, this target can be rescued efficiently from genomic DNA prepared from tissues of the treated mice using restriction minus, in vitro lambda phage packaging extract and restriction minus <em>Escherichia</em> <em>coli</em> plating cultures. Mutations in the target gene appear as colorless plaques on a background of blue plaques when plated on indicator agar. Spontaneous background levels were ′1 × 10<sup>−5</sup> in each of three mouse lineages analyzed. Exposure of lambda transgenic mice to <em>N</em>-ethyl-<em>N</em>-nitrosourea resulted in as much as a 14-fold induction in detected mutations over background levels. The assay is currently being modified to incorporate <em>lac</em>I as the target for ease of mutation detection as well as in vivo excision properties of the Lambda ZAP vector, facilitating sequence analysis of mutant plaques.</p></div>","PeriodicalId":77714,"journal":{"name":"Gene analysis techniques","volume":"7 8","pages":"Pages 212-218"},"PeriodicalIF":0.0,"publicationDate":"1990-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0735-0651(90)90003-X","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"13305243","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 79
Expression of animal and human retroviral gene products in Escherichia coli with the λPL promoter pJL6 vector system 用λPL启动子pJL6载体系统在大肠杆菌中表达动物和人逆转录病毒基因产物
Gene analysis techniques Pub Date : 1990-11-01 DOI: 10.1016/0735-0651(90)90023-9
Kenneth P. Samuel , Richard Ascione , Stavros D. Kottaridis , Arun K. Seth , James A. Lautenberger , Mohammed Zuber , John Strouboulis , Takis S. Papas
{"title":"Expression of animal and human retroviral gene products in Escherichia coli with the λPL promoter pJL6 vector system","authors":"Kenneth P. Samuel ,&nbsp;Richard Ascione ,&nbsp;Stavros D. Kottaridis ,&nbsp;Arun K. Seth ,&nbsp;James A. Lautenberger ,&nbsp;Mohammed Zuber ,&nbsp;John Strouboulis ,&nbsp;Takis S. Papas","doi":"10.1016/0735-0651(90)90023-9","DOIUrl":"https://doi.org/10.1016/0735-0651(90)90023-9","url":null,"abstract":"","PeriodicalId":77714,"journal":{"name":"Gene analysis techniques","volume":"7 7","pages":"Pages 178-208"},"PeriodicalIF":0.0,"publicationDate":"1990-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0735-0651(90)90023-9","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71863202","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 2
Simultaneous isolation of total cellular RNA and DNA from tissue culture cells using phenol and lithium chloride 用苯酚和氯化锂同时从组织培养细胞中分离总细胞RNA和DNA
Gene analysis techniques Pub Date : 1990-11-01 DOI: 10.1016/0735-0651(90)90022-8
Sandeep Raha , Frank Merante , Gerald Proteau , Juta K. Reed
{"title":"Simultaneous isolation of total cellular RNA and DNA from tissue culture cells using phenol and lithium chloride","authors":"Sandeep Raha ,&nbsp;Frank Merante ,&nbsp;Gerald Proteau ,&nbsp;Juta K. Reed","doi":"10.1016/0735-0651(90)90022-8","DOIUrl":"10.1016/0735-0651(90)90022-8","url":null,"abstract":"<div><p>A rapid procedure for the isolation of intact total cellular RNA from cultured cells is described. This method combines the simultaneous disruption of cells and extraction of nucleic acids in a single step with the use of phenol and a buffer containing 100 mM LiCl. Total cellular RNA can be isolated in approximately 2 hours. The yield and quality of the RNA is comparable to the more widely employed methods requiring extensive preparatory steps such as extraction using guanidinium thiocyanate and subsequent CsCl gradient centrifugation. The RNA isolated using our procedure contains transcripts up to 10 kb in length and is suitable for Northern analysis. This procedure also yields high-molecular-weight DNA, which is a suitable substrate for restriction endonucleases.</p></div>","PeriodicalId":77714,"journal":{"name":"Gene analysis techniques","volume":"7 7","pages":"Pages 173-177"},"PeriodicalIF":0.0,"publicationDate":"1990-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0735-0651(90)90022-8","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"12870371","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 35
Magnetic DNA hybridization properties of oligonucleotide probes attached to superparamagnetic beads and their use in the isolation of poly(A) mRNA from eukaryotic cells 超顺磁珠寡核苷酸探针的磁DNA杂交特性及其在真核细胞中聚(A) mRNA分离中的应用
Gene analysis techniques Pub Date : 1990-10-01 DOI: 10.1016/0735-0651(90)90028-E
Erik Hornes, Lars Korsnes
{"title":"Magnetic DNA hybridization properties of oligonucleotide probes attached to superparamagnetic beads and their use in the isolation of poly(A) mRNA from eukaryotic cells","authors":"Erik Hornes,&nbsp;Lars Korsnes","doi":"10.1016/0735-0651(90)90028-E","DOIUrl":"10.1016/0735-0651(90)90028-E","url":null,"abstract":"<div><p>Poly(A) messenger RNA is generally purified from total RNA using oligo(dT) cellulose affinity chromatography or centrifugation through spin columns. We present a new method for rapid purification of poly(A) mRNA using oligo(dT) probes attached to superparamagnetic beads. By magnetic separation, washing, and elution, pure mRNA is obtained from living cells within 10 minutes. This procedure works for crude RNA preparations or cell lysates that would otherwise clog standard oligo(dT) cellulose column systems. The present method reduces the risk of degradation, is highly efficient, and can easily be scaled up or down.</p></div>","PeriodicalId":77714,"journal":{"name":"Gene analysis techniques","volume":"7 6","pages":"Pages 145-150"},"PeriodicalIF":0.0,"publicationDate":"1990-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0735-0651(90)90028-E","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"13232245","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 47
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