Abstract:
:A solid-phase method is described to create subclones, suitable for DNA sequencing, from lambda or cosmid libraries. The purified target DNA is sonicated and two linkers, with one oligonucleotide biotinylated in the 5'-end, are ligated to the ends of the fragments produced by sonication. After size separation, the fragments are immobilised onto a solid support and the non-biotinylated strand of each immobilised fragment is eluted. In this way, a library of single-stranded fragments is obtained. All fragments contain 'universal' flanking sequences of 22 bases introduced by the linker ligation. These flanking sequences can subsequently be used for solid-phase cloning into a single-stranded vector containing the complementary sequences. Thus, cloning can be achieved without the use of ligase or restriction enzymes. The resulting subclones are used for direct solid-phase sequencing and the immobilised strand can be used to selectively remove homologous DNA from the library of single-stranded fragments. Thus, a sublibrary of non-sequenced fragments can be created. Here, we show that a library of clones, suitable for direct solid-phase sequencing, can be obtained starting with lambda DNA. The efficiency of selective hybridisation of homologous and non-homologous fragments was investigated. The possibility of using this approach for automated cloning strategies for large-scale genomic and cDNA sequencing is discussed.
journal_name
J Biotechnoljournal_title
Journal of biotechnologyauthors
Hultman T,Uhlén Mdoi
10.1016/0168-1656(94)90038-8subject
Has Abstractpub_date
1994-06-30 00:00:00pages
229-38issue
2-3eissn
0168-1656issn
1873-4863pii
0168-1656(94)90038-8journal_volume
35pub_type
杂志文章abstract::This study demonstrates the applicability of pressurized stirred tank bioreactors for oxygen transfer enhancement in aerobic cultivation processes. The specific power input and the reactor pressure was employed as process variable. As model organism Escherichia coli, Arxula adeninivorans, Saccharomyces cerevisiae and ...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2007.06.010
更新日期:2007-10-31 00:00:00
abstract::Whole metagenome shotgun sequencing provides information about the gene content and the composition of microbial communities provided that the processing of the samples does not introduce a methodology-driven bias. We tested the impact of DNA isolation and storage period on the metagenome profile. Deep throat swabs we...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2017.01.001
更新日期:2017-05-20 00:00:00
abstract::A control strategy to avoid development of synchronous growth in carbohydrate limited Saccharomyces cerevisiae cultivations is proposed and experimentally investigated. The basic idea is to control the metabolic flux through the pathways by manipulating the substrate feed rate to keep the ethanol concentration at a lo...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/s0168-1656(97)01662-3
更新日期:1997-02-28 00:00:00
abstract::Pseudomonas alcaligenes secretes a lipase with a high pH optimum, which has interesting properties for application in detergents. The expression of the lipase is strongly dependent on the presence of lipids in the growth medium such as soybean oil. The promoter of the gene was characterized and found to have resemblan...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/s0168-1656(00)00397-7
更新日期:2001-03-09 00:00:00
abstract::This paper introduces a novel approach to enhance the strains' biodegradation of 4-chloronitrobenzene by utilizing the synergistic effect of the organic reductant mannitol and the substrate beef extraction. Our results demonstrate that 4-chloronitrobenzene could not be an available nitrogen source to support target st...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2017.10.005
更新日期:2017-12-20 00:00:00
abstract::Much attention has been paid to membrane bioreactors, especially to the newly developed charged membrane bioreactor (CMBR) for recycling the native form of the coenzyme NAD(P)H. Charged membranes with anionic groups effectively retained the free nicotinamide coenzyme due to the electrostatic repulsion between membrane...
journal_title:Journal of biotechnology
pub_type: 杂志文章,评审
doi:10.1016/0168-1656(90)90010-9
更新日期:1990-05-01 00:00:00
abstract::The feed profile of glucose during fedbatch cultivation could be used to influence the retention of the periplasmic product ZZ-cutinase. An increased feed rate led to a higher production rate but also to an increased specific leakage, which reduced the periplasmic retention. Three growth rates: 0.3, 0.2 and 0.1 h(-1) ...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2008.05.002
更新日期:2008-07-31 00:00:00
abstract::We cloned and characterized a preprochymosin cDNA from the abomasum of milk-fed kid goats. This cDNA contained an open reading frame that predicts a polypeptide of 381 amino acid residues, with a signal peptide and a proenzyme region of 16 and 42 amino acids, respectively. Comparison of the caprine preprochymosin sequ...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2004.06.002
更新日期:2004-10-19 00:00:00
abstract::The importance of RNA-based regulation is becoming more and more evident. Genome-wide sequencing efforts have shown that the majority of the DNA in eukaryotic genomes is transcribed. Advanced high-throughput techniques like CLIP for the genome-wide detection of RNA-protein interactions have shown that post-transcripti...
journal_title:Journal of biotechnology
pub_type: 杂志文章,评审
doi:10.1016/j.jbiotec.2017.05.019
更新日期:2017-11-10 00:00:00
abstract::Planococcus rifietoensis M8(T) (=DSM 15069(T)=ATCC BAA-790(T)) is a halotolerant bacterium with potential plant growth promoting properties isolated from an algal mat collected from a sulfurous spring in Campania (Italy). This paper presents the first complete genome of P. rifietoensis M8(T). Genes coding for various ...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2016.01.026
更新日期:2016-03-10 00:00:00
abstract::The large scale production of a monoclonal anti-progesterone antibody in serum free medium followed by affinity chromatography on protein G lead to a contamination of the antibody sample with a protein of about 14 kDa. This protein was identified by mass spectrometry as secretory leukocyte protease inhibitor (SLPI). T...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2011.12.025
更新日期:2012-03-31 00:00:00
abstract::In this paper we describe the antigen recognition characteristics, variable region base and amino acid sequence, and performance as immunoaffinity chromatography ligands of two MAb specific to the alpha determinant of the HBsAg, derived from the same fusion. We show that the epitope recognized by CB-Hep.0 (IgM) is pro...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/s0168-1656(97)00103-x
更新日期:1997-08-11 00:00:00
abstract::The genus Enterococcus occurs as native microbiota of fermented products due to its broad environmental distribution and its resistance to salt concentrations. Enterococcus faecalis F, a non-pathogenic strain isolated from a ripened cheese, has demonstrated useful enzymatic capabilities, a probiotic behavior and antib...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2018.06.346
更新日期:2018-10-10 00:00:00
abstract::Oxygen is a key substrate in animal cell metabolism. It has been reported that the oxygen uptake rate (OUR) is a good indicator of cellular activity, and even under some conditions, a good indicator of the number of viable cells. The measurement of OUR is difficult due to many different reasons. In particular, the ver...
journal_title:Journal of biotechnology
pub_type: 杂志文章,评审
doi:10.1016/s0168-1656(98)00046-7
更新日期:1998-08-12 00:00:00
abstract::Five nominally freshwater microalgae (Chlorella vulgaris, Choricystis minor, Neochloris sp., Pseudococcomyxa simplex, Scenedesmus sp.) with a known ability to produce high-levels of lipids for possible use as fuel oils were evaluated for their ability to thrive and produce lipids in seawater and brackish water. Only C...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2016.11.029
更新日期:2017-09-10 00:00:00
abstract::The application of biocalorimetry to fermentation processes offers advantageous insights, while being less complex compared to other, sophisticated PAT solutions. Although the general concept is established, calorimetric methods vary in detail. In this work, a special approach, called heat compensation calorimetry, wa...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2017.12.002
更新日期:2018-01-20 00:00:00
abstract::Primary human lymphocytes and macrophages are an important target cells for human immunodeficiency virus (HIV). For targeted gene transfer into CD4(+) lymphocytes and macrophages, we constructed HIV vectors with envelope glycoprotein (gp120) from the T-cell tropic BH10 strain and the macrophage tropic SF162, and devel...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2007.01.004
更新日期:2007-05-01 00:00:00
abstract::A simple and accurate analysis system for the estimation of recombination efficiency in vivo using fluorescence-activated cell sorting (FACS) was designed and was subsequently used to compare the efficiency of recombination related to different spacer mutants. F(3) and F(5) mutant sequences were used for Flpe-mediated...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2006.07.027
更新日期:2007-01-10 00:00:00
abstract::Biosurfactants are surface-active agents produced by microorganisms and show increasing significance in various industrial applications. A great variety of these secondary metabolites are described to occur within actinomycetes, amongst trehalose lipids and oligosaccharide lipids produced by the family Tsukamurellacea...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2015.06.424
更新日期:2015-10-10 00:00:00
abstract::In this paper, single-stranded (ss)DNA aptamers with capability to distinguish differentiated PC12 cells from normal PC12 cells were selected by subtractive systematic evolution of ligands by exponential enrichment (SELEX) method. Before each round of selection, randomized ssDNAs were incubated with regular PC12 cells...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/s0168-1656(02)00360-7
更新日期:2003-04-10 00:00:00
abstract::The conversion of olive mill wastewater (OMW) into high added value lipids containing polyunsaturated fatty acids (PUFA), in parallel with a significant phenolic removal by selected strains of Zygomycetes, is reported here for the first time. The growth of Mortierella isabellina, Mortierella ramanniana, Cunninghamella...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2013.11.015
更新日期:2014-01-20 00:00:00
abstract::The adsorption behaviors of two glucanases, TvEG and TrCel7A, on three lignins were investigated. Three lignins were isolated from raw aspen and its pretreated solid residue. The isolated lignins were labeled as Asp-MWL, DA-MWL (pretreated by dilute acid), and GL-MWL (pretreated by green liquor), respectively. The sur...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2020.07.013
更新日期:2020-11-10 00:00:00
abstract::New antifungal compounds that circumvent the resistance of the pathogen by directly damaging yeast cell surface structures are promising agents for the treatment of fungal infections, due to their different mechanism of action from current clinically used antifungal drugs. We present here a rapid and cost-effective fl...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2016.06.023
更新日期:2016-09-10 00:00:00
abstract::For biotechnological production of coenzyme Q(10) (CoQ(10)) in recombinant Escherichia coli, three genetic manipulations were performed: heterologous expression of decaprenyl diphosphate synthase (Dps) from Agrobacterium tumefaciens, deletion of endogenous octaprenyl diphosphate synthase (IspB), and overexpression of ...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2009.04.010
更新日期:2009-10-12 00:00:00
abstract::The polysaccharide xanthan which is produced by the γ-proteobacterium Xanthomonas campestris is used as a food thickening agent and rheologic modifier in numerous food, cosmetics and technical applications. Its great commercial importance stimulated biotechnological approaches to optimize the xanthan production. By ta...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2016.11.032
更新日期:2017-09-10 00:00:00
abstract::Bacillus velezensis G341 can suppress plant pathogens by producing antagonistic active compounds including bacillomycin D, fengycin, and (oxy) difficidin. The complete genome sequence of this bacterium was characterized by one circular chromosome of 4,009,746bp with 3953 open reading frames. The genome contained 36 ps...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2015.07.005
更新日期:2015-10-10 00:00:00
abstract::The effects of a deletion of the amtR gene, encoding the master regulator of nitrogen control in Corynebacterium glutamicum, were investigated by metabolome and transcriptome analyses. Compared to the wild type, different metabolite patterns were observed in respect to glycolysis, pentose phosphate pathway, citric aci...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2008.10.009
更新日期:2009-03-10 00:00:00
abstract::Bacillus thuringiensis is a ubiquitous spore-forming bacterium and has been widely used as a biopesticide for controlling agricultural insects by the production of insecticidal crystal proteins (ICPs). B. thuringiensis YBT-1518 displays effective toxicity to nematodes. This strain harbors three nematicidal crystal pro...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2013.11.023
更新日期:2014-02-10 00:00:00
abstract::Chimeric virus-like particles (VLP) are known as promising tools in the development of safe and effective subunit vaccines. Recently, a technology platform to produce VLP based on the small surface protein (dS) of the duck hepatitis B virus was established. In this study, chimeric VLP were investigated displaying the ...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2019.10.008
更新日期:2019-12-20 00:00:00
abstract::The poly(beta-hydroxybutyrate) (PHB) biosynthetic genes of Ralstonia eutropha that are organized in a single operon (phaCAB) have been cloned in Escherichia coli, where the expression of the genes in the wild-type pha operon from plasmid pTZ18U-PHB leads to the formation of 50-80% PHB/celldry mass when the cells are g...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/s0168-1656(98)00127-8
更新日期:1998-10-27 00:00:00