Abstract:
:Identification of expressed sequence tags (ESTs) in large genomic segments is an important step in positional cloning and genomic mapping studies. A simple and efficient polymerase chain reaction (PCR)-based approach is described here to identify coding sequences in large genomic fragments of DNA cloned in vectors such as yeast artificial chromosome (YAC) vectors. The method is based on blocking of sequences such as repetitive and GC rich sequences in the genomic DNA immobilized on nylon paper discs prior to hybridization of the discs to cDNA library, and recovery of the selected cDNAs by the PCR. Single or multiple cDNA libraries can be used in the selection procedure. The procedure has been used successfully also with total yeast DNA containing a YAC.
journal_name
Mol Biotechnoljournal_title
Molecular biotechnologyauthors
Parimoo Sdoi
10.1007/BF02760779subject
Has Abstractpub_date
1997-12-01 00:00:00pages
255-68issue
3eissn
1073-6085issn
1559-0305journal_volume
8pub_type
杂志文章abstract::The major online single nucleotide polymorphism (SNP) databases freely available as research tools for genetic analysis are explained, reviewed, and compared. An outline is given of the search strategies that can be used with the most extensive current SNP databases: National Centre for Biotechnology Information (NCBI...
journal_title:Molecular biotechnology
pub_type: 杂志文章,评审
doi:10.1385/mb:35:1:65
更新日期:2007-01-01 00:00:00
abstract::In this paper, we report cloning of a pectate lyase gene from Bacillus amyloliquefaciens S6 (pelS6), and biochemical characterization of the recombinant pectate lyase. PelS6 was found to be identical with B. subtilis 168 pel enzyme with 100% amino acid sequence homology. Although these two are genetically very close, ...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-019-00194-2
更新日期:2019-09-01 00:00:00
abstract::A single-stage polymerase-based procedure is described that allows extensive modifications of DNA. The version described here uses the QuikChange Site-Directed Mutagenesis System kit supplied by Stratagene. The original protocol is replaced by a single-stage method in which linear production of complementary strands i...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:29:3:225
更新日期:2005-03-01 00:00:00
abstract::The induction of several secondary metabolites in plants is one of the most commonly observed effects after the external addition of methyl jasmonate (MeJA). After the elicitation of Catharanthus roseus hairy roots with different concentrations of MeJA, changes in the accumulation of alkaloids such as ajmalicine, serp...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-008-9111-2
更新日期:2009-03-01 00:00:00
abstract::RNA editing is one of the post-transcriptional RNA processes. RNA editing generates RNA and protein diversity in eukaryotes and results in specific amino acid substitutions, deletions, and changes in gene expression levels. Adenosine-to-inosine RNA editing represents the most important class of editing in human and af...
journal_title:Molecular biotechnology
pub_type: 杂志文章,评审
doi:10.1007/s12033-012-9498-7
更新日期:2012-09-01 00:00:00
abstract::Production of monoclonal antibodies and pharmaceutical proteins in transgenic plants has been the focus of many research efforts for close to 30 years. Use of plants as bioreactors reduces large-scale production costs and minimizes risk for human pathogens contamination. Stable nuclear transformation of the plant geno...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-018-0075-6
更新日期:2018-06-01 00:00:00
abstract::Monoclonal antibodies (mAbs) are the fastest growing class of biopharmaceuticals reflecting their diverse applications in research and the clinic. The correct glycosylation of mAbs is required to elicit effector functions such as complement-dependent and antibody-dependent cell-mediated cytotoxicity, although these ma...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-012-9531-x
更新日期:2013-03-01 00:00:00
abstract::The gram-negative anaerobic oral bacterium Porphyromonas gingivalis initiates periodontal disease through fimbrial attachment to saliva-coated oral surfaces. To study the effects of immunomodulation on enhancement of subunit vaccination, the expression in E. coli and immunogenicity of P. gingivalis fimbrial protein (F...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-008-9102-3
更新日期:2009-02-01 00:00:00
abstract::As the use of adenoviral vectors in gene therapy protocols increases, there is a corresponding need for rapid, accurate, and reproducible titer methods. Multiple methods currently exist for determining titers of recombinant adenoviral vector, including optical absorbance, electron microscopy, fluorescent focus assay, ...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:14:3:197
更新日期:2000-03-01 00:00:00
abstract::In this study, we investigated the relative expression of the Rous sarcoma virus (RSV) promoter-driven expression of enhanced green fluorescent protein (EGFP) in fibroblasts of transgenic quails. We analyzed the direct influence of CpG methylation of the RSV promoter on the transcriptional activity of delivered transg...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-011-9393-7
更新日期:2011-11-01 00:00:00
abstract::Pasteurella multocida is the main cause of haemorrhagic septicaemia (HS) outbreak in livestock, such as cattle and buffaloes. Conventional vaccines such as alum-precipitated or oil-adjuvant broth bacterins were injected subcutaneously to provide protection against HS. However, the immunity developed is only for short ...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-020-00244-0
更新日期:2020-05-01 00:00:00
abstract::Jet-injection has become an applicable technology among other established nonviral delivery systems, such as particle bombardment or in vivo electroporation. The low-volume jet injector employed in this study uses compressed air to inject solutions of 1.5-10 microL containing naked DNA into the desired tissue. The nov...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:21:2:105
更新日期:2002-06-01 00:00:00
abstract::Ageing is a complex and progressive phenomenon, during which the accumulation of morphological and chemical changes seriously compromises the capacity of the cells to proliferate and fulfil their biological tasks. The increase in the average age of the world population, associated with a higher occurrence of age-relat...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-019-00154-w
更新日期:2019-03-01 00:00:00
abstract::ATP-binding cassette transporter G2 (ABCG2) gene encodes a protein that has a wide variety of substrates and is responsible for the active secretion of clinically and toxicologically important molecules into milk. Although known in many species, this marks the first time this gene product has been reported in goats. I...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-007-9024-5
更新日期:2008-05-01 00:00:00
abstract::Plants synthesize amino acids by collateral metabolic pathways using primary elements carbon and oxygen from air, hydrogen from water in soil and nitrogen from soil. Following synthesis, amino acids are immediately used for metabolism, transient storage or transported to the phloem. Different families of transporters ...
journal_title:Molecular biotechnology
pub_type: 杂志文章,收录出版
doi:10.1007/s12033-020-00240-4
更新日期:2020-03-01 00:00:00
abstract::Alternaria blight caused by Alternaria brassicicola (Schwein.) Wiltshire and A. brassicae (Berk.) Sacc., is one of the most important disease of rapeseed-mustard, characterized by the formation of spots on leaves, stem, and siliquae with premature defoliation and stunting of growth. These two species are very difficul...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-014-9784-7
更新日期:2014-11-01 00:00:00
abstract::Terminal Restriction Fragment Length Polymorphism (T-RFLP) or Fluorescent Polymerase Chain Reaction/Restriction Fragment Length Polymorphism (FluRFLP) have made a significant impact on the way in which PCR products amplified from mixed community DNA extracts have been assessed. Technically, these approaches are essent...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:16:3:261
更新日期:2000-11-01 00:00:00
abstract::A computer program (PCBI) was developed to quickly calculate codon bias index (CBI). PCBI can analyze a gene containing introns. The 22 preferred codons defined from Saccharomyces cerevisiae were used in PCBI as the standard to measure the CBI values. However, users can modify the preferred codons to suit each organis...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/BF02760858
更新日期:1998-10-01 00:00:00
abstract::A bacterial artificial chromosome library of the causal agent of the Black Sigatoka leaf spot disease of banana and plantain, Mycosphaerella fijiensis, has been constructed using a non-sphaeroplasting technique and characterized using both homologous and heterologous probes. After first and a second size selection of ...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-007-0016-2
更新日期:2007-05-01 00:00:00
abstract::High-quality RNA preparations are critical for further applications such as reverse transcriptase-polymerase chain reaction (RT-PCR) transcript amplifications, and elaboration of cDNA and expressed sequence tag libraries. Melanins are phenolic compounds present in many fungi and apparently play key roles in fungi path...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:34:1:45
更新日期:2006-09-01 00:00:00
abstract::While conducting our research on mutations in the human blood platelet glycoprotein Ib-alpha (GPIbalpha) gene, we detected an unusual deletion of 84 bp. This deletion took place in vitro, during PCR and between two direct repeats. It was observed that the deletion could be detected either by the direct sequencing of t...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-008-9059-2
更新日期:2008-09-01 00:00:00
abstract::A sensitive method for staining proteins after transfer from polyacrylamide gels to nitrocellulose paper is described. Transferred proteins are first derivatized by reaction of the nitrocellulose replica with sulfosuccinimidobiotin and are then reacted sequentially with streptavidin, rabbit anti-streptavidin, and hors...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:15:2:161
更新日期:2000-06-01 00:00:00
abstract::Dye-based protein determination assays are widely used to estimate protein concentration, however various reports suggest that the response is dependent on the composition and sequence of the protein, limiting confidence in the resulting concentration estimates. In this study a diverse set of model proteins representi...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-007-0038-9
更新日期:2007-10-01 00:00:00
abstract::Host protein synthesis is shut down in the lytic baculovirus expression vector system (BEVS). This also affects host proteins involved in routing secretory proteins through the endoplasmic reticulum (ER)-Golgi system. It has been demonstrated that a secretory alkaline phosphatase-EGFP fusion protein (SEFP) can act as ...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-012-9545-4
更新日期:2013-05-01 00:00:00
abstract::Hereditary cancers account for approximately 10 % of breast and ovarian cancers. Mutations of the BRCA1 and BRCA2 genes, encoding two proteins involved in DNA repair, underlie most cases of such hereditary cancers. Women with BRCA mutations develop breast cancer in 50-80 % of cases and ovarian cancer in 10-40 % of cas...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-012-9646-0
更新日期:2013-07-01 00:00:00
abstract::Modern sugarcane cultivars have complex genetic characteristics and low fertility that render their genetic improvement through traditional breeding difficult. Genetic engineering methodology to introduce foreign genes provides new opportunities for the genetic improvement of sugarcane cultivars. One of prerequisites ...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:28:2:113
更新日期:2004-10-01 00:00:00
abstract::Transgenic rice seed can be utilized as a bioreactor to produce high-value recombinant proteins. Mouse interleukin 4 (mIL-4) and mIL-6 were specifically expressed as secretory proteins in rice endosperm by ligating the N-terminal glutelin B-1 (GluB-1) signal peptide and the C-terminal KDEL endoplasmic reticulum retent...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-016-9920-7
更新日期:2016-04-01 00:00:00
abstract::The term "coincidence cloning" encompasses a wide range of methodologies, the aim of which is to isolate DNA sequences which occur in both of two input DNA sources. The nature of these input DNAs may be genomic or cDNA, cloned or uncloned, and as such the far reaching applicability of the techniques can be imagined. I...
journal_title:Molecular biotechnology
pub_type: 杂志文章,评审
doi:10.1007/BF02900362
更新日期:1996-06-01 00:00:00
abstract::Monoclonal antibodies (Mabs) against human chorionic gonadotropin hormone (hCG) were raised by hybridoma technology using Sp2/0 myeloma cells as fusion partner. Sixty-five percent of the total culture wells exhibited hybrid growth and 8% of the total wells (13 culture wells) contained anti-hCG secreting hybrids. A pos...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:17:2:119
更新日期:2001-02-01 00:00:00
abstract::Cyclooxygenase (COX)-2 plays an important role in the development of cancer and has been recognized as a potential therapeutic target. Because nonsteroidal anti-inflammatory drugs (NSAIDs) are able to inhibit the activity of this enzyme, the potential efficacy of such drugs for purposes of cancer prevention or therapy...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:34:3:329
更新日期:2006-11-01 00:00:00