Genetic analysis, techniques and applications最新文献_第7页

Extremely-rapid RNA detection in dot blots with digoxigenin-labeled RNA probes 用地高辛标记的RNA探针进行点印迹的极快速RNA检测

Genetic analysis, techniques and applications Pub Date : 1998-10-01 DOI: 10.1016/S1050-3862(98)00003-5

Chih-Yang Huang, Matsuo Kasai, Dennis E. Buetow

引用次数: 10

A new technique of screening gene mutation—fluorescent fingerprinting 一种筛选基因突变的新技术——荧光指纹

Genetic analysis, techniques and applications Pub Date : 1998-10-01 DOI: 10.1016/S1050-3862(98)00008-4

Jingzhong Liu , Hua Xiang , Yan Zhou , Zhangling Zhu , Jing Zhang

引用次数: 3

Construction and sequencing of new derivatives of the pIN-III-ompA secretion vector pIN-III-ompA分泌载体新衍生物的构建与测序

Genetic analysis, techniques and applications Pub Date : 1998-10-01 DOI: 10.1016/S1050-3862(98)00005-9

Alan Fauconnier, Ary Van Elsen, Alex Bollen

引用次数: 5

Trans-kingdom conjugation offers a powerful gene targeting 跨界结合提供了一种强大的基因靶向

Genetic analysis, techniques and applications Pub Date : 1998-01-01 DOI: 10.1016/S1050-3862(97)10003-1

Masanobu Nishikawa , Kazuo Yoshida

引用次数: 12

Cloning, sequence analysis and expression in E. coli of the DNA polymerase I gene from Chloroflexus aurantiacus, a green 绿色植物金菖蒲DNA聚合酶I基因的克隆、序列分析及在大肠杆菌中的表达

Genetic analysis, techniques and applications Pub Date : 1998-01-01 DOI: 10.1016/S1050-3862(97)10002-X

Marianne Tvermyr , Bjørn E Kristiansen, Tom Kristensen

{"title":"Cloning, sequence analysis and expression in E. coli of the DNA polymerase I gene from Chloroflexus aurantiacus, a green","authors":"Marianne Tvermyr , Bjørn E Kristiansen, Tom Kristensen","doi":"10.1016/S1050-3862(97)10002-X","DOIUrl":"10.1016/S1050-3862(97)10002-X","url":null,"abstract":"<div>We have cloned and sequenced the polA gene from Chloroflexus aurantiacus, a green nonsulfur eubacterium, and expressed the recombinant protein in Escherichia coli. One open reading frame encodes a protein with 942 amino acids showing 38% identity with DNA polymerase I from E. coli. Sequence alignments with other members of DNA polymerase family A and analysis of the separate domains show that the central 3′-5′ exonuclease domain is 30% identical to the corresponding E. coli domain and that three sequence motifs associated with 3′-5′ exonuclease activity are conserved. Also, a protein fraction from E. coli expressing the Chloroflexus polymerase contains a thermostable 3′-5′ exonucleolytic activity, indicating that this activity is present in the enzyme, in agreement with the sequence analysis. The N-terminal 5′-3′ exonuclease domain and the C-terminal polymerase domain show 31 and 46% identity, respectively, with the corresponding E. coli domains and all sequence motifs associated with these two enzymatic activities also are conserved. Since several DNA polymerase I enzymes lack the proofreading activity associated with the central domain it has been suggested that the ancestral polA gene contained only the two more conserved N- and C-terminal domains and that the proofreading 3′-5′ exonuclease domain was introduced later in those eubacterial branches that have this activity. Our data indicate a different scenario where the ancestral polA gene contained both the exonucleolytic activities in addition to the polymerase activity and where several eubacterial branches lost the polymerase-associated proofreading activity during evolution.</div>","PeriodicalId":77142,"journal":{"name":"Genetic analysis, techniques and applications","volume":"14 3","pages":"Pages 75-83"},"PeriodicalIF":0.0,"publicationDate":"1998-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S1050-3862(97)10002-X","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20449105","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: 4

Assay of transfection rate in insect cells on a single cell level 单细胞水平上昆虫细胞转染率测定

Genetic analysis, techniques and applications Pub Date : 1998-01-01 DOI: 10.1016/S1050-3862(97)10001-8

Bernd Rautenstrauss , Christina Fuchs , Arif Ekici , Eva Nelis , Christine Van Broeckhoven , Thomas Liehr

引用次数: 6

ZFX and ZFY loci in water buffalo (Bubalus bubalis): 水牛(Bubalus bubalis) ZFX和ZFY位点:

Genetic analysis, techniques and applications Pub Date : 1998-01-01 DOI: 10.1016/S1050-3862(97)10004-3

Anita Pande, S.M Totey

引用次数: 7

Application of nested PCR and mass spectrometry for DNA-based virus detection: HBV-DNA detected in the majority of isolated anti-HBc positive sera 巢式PCR和质谱法在基于dna的病毒检测中的应用:在大多数分离的抗hbc阳性血清中检测到HBV-DNA

Genetic analysis, techniques and applications Pub Date : 1998-01-01 DOI: 10.1016/S1050-3862(97)10006-7

Christian Jurinke , Bernhard Zöllner , Heinz-Hubert Feucht , Dirk van den Boom , Anette Jacob , Susanne Polywka , Rainer Laufs , Hubert Köster

{"title":"Application of nested PCR and mass spectrometry for DNA-based virus detection: HBV-DNA detected in the majority of isolated anti-HBc positive sera","authors":"Christian Jurinke , Bernhard Zöllner , Heinz-Hubert Feucht , Dirk van den Boom , Anette Jacob , Susanne Polywka , Rainer Laufs , Hubert Köster","doi":"10.1016/S1050-3862(97)10006-7","DOIUrl":"10.1016/S1050-3862(97)10006-7","url":null,"abstract":"<div>DNA preparations from three different groups of serum samples were examined for HBV-DNA via a nested polymerase chain reaction assay (lower detection limit: 10 viral genomes in 100 μl serum): Group I consisted of 11 uninfected control sera, group II consisted of sera obtained from 11 HBV infected patients and group III consisted of 21 isolated anti-HBc positive samples. The 21 samples from group III were HBV-DNA negative according to a conventional non-nested PCR assay and hybridization with a 32P-labelled probe. Using nested PCR and mass spectrometry, HBV-DNA was detected in none of group I and in all of group II samples. In 11 out of 21 (52%) of the isolated anti-HBc positive sera from group III, HBV-DNA was detected. No correlation was observed between HBV-DNA positivity and anti-HBc titers. Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry provided a fast, sensitive and non-radioactive assay for the detection of PCR products without the need for gel electrophoresis or hybridization with labelled probes.</div>","PeriodicalId":77142,"journal":{"name":"Genetic analysis, techniques and applications","volume":"14 3","pages":"Pages 97-102"},"PeriodicalIF":0.0,"publicationDate":"1998-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S1050-3862(97)10006-7","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20449108","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: 21

Cre/loxP-mediated excision and amplification of large segments of the Escherichia coli genome Cre/ loxp介导的大肠杆菌基因组大片段的切除和扩增

Genetic analysis, techniques and applications Pub Date : 1998-01-01 DOI: 10.1016/S1050-3862(97)10005-5

Young G. Yoon, Ju H. Cho, Sun C. Kim

{"title":"Cre/loxP-mediated excision and amplification of large segments of the Escherichia coli genome","authors":"Young G. Yoon, Ju H. Cho, Sun C. Kim","doi":"10.1016/S1050-3862(97)10005-5","DOIUrl":"10.1016/S1050-3862(97)10005-5","url":null,"abstract":"<div>The isolation and amplification of large, predetermined segments of a genome from its host have been explored. The prototype of our approach was the excisional replication of some viruses such as the λ-lysogen. Similar machinery was used to excise and amplify large genomic segments of Escherichia coli in its host. Two loxP sequences for a site-specific recombinase Cre, together with a conditional replication origin (π-dependent γ-ori), were inserted into the genome by homologous recombination at predetermined sites, 50–100 kb apart. Cre and pir200 which encodes the site-specific recombinase Cre and an ori-specific replication protein π, respectively, were also introduced into the genome. The predetermined genomic segments flanked by the loxP sequences were excised and amplified upon induction of the cre and pir200 genes which were under the control of the tet promoter. This excised and amplified DNA could be easily purified as a large plasmid. This procedure can provide an alternative to conventional cloning methods by obtaining predetermined large genomic segments directly from the original organisms. In this study, using the Cre/loxP site-specific recombination and π/γ-ori replication system of plasmid R6K, a procedure was devised that could isolate a large segment of the E. coli genome and demonstrated the feasibility of the procedure by excising and amplifying the 50-kb trg-narZ and 100-kb trg-hipA regions of the E. coli W3110 genome.</div>","PeriodicalId":77142,"journal":{"name":"Genetic analysis, techniques and applications","volume":"14 3","pages":"Pages 89-95"},"PeriodicalIF":0.0,"publicationDate":"1998-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S1050-3862(97)10005-5","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20449107","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: 21

Rapid method for detection of point mutations using mismatch binding protein (MutS) and an optical biosensor 利用错配结合蛋白和光学生物传感器快速检测点突变的方法

Genetic analysis, techniques and applications Pub Date : 1997-07-01 DOI: 10.1016/S1050-3862(97)00009-0

Masanori Gotoh, Masahisa Hasebe, Tomoko Ohira, Yukio Hasegawa, Yasuro Shinohara, Hiroyuki Sota, Junji Nakao, Mariko Tosu

引用次数: 34