Abstract:
BACKGROUND:Social amoeba, Dictyostelium discoideum, is a well-established model organism for studying cellular physiology and developmental pattern formation. Its haploid genome facilitates functional analysis of genes by a single round of mutagenesis including targeted disruption. Although the efficient generation of knockout strains based on an intrinsically high homologous recombination rate has been demonstrated, successful reports for knockin strains have been limited. As social amoeba has an exceptionally high adenine and thymine (A/T)-content, conventional plasmid-based vector construction has been constrained due to deleterious deletion in E. coli. RESULTS:We describe here a simple and efficient strategy to construct GFP-knockin cassettes by using a linear DNA cloning vector derived from N15 bacteriophage. This allows reliable handling of DNA fragments whose A/T-content may be as high as 85 %, and which cannot be cloned into a circular plasmid. By optimizing the length of recombination arms, we successfully generate GFP-knockin strains for five genes involved in cAMP signalling, including a triple-colour knockin strain. CONCLUSIONS:This robust strategy would be useful in handling DNA fragments with biased A/T-contents such as the genome of lower organisms and the promoter/terminator regions of higher organisms.
journal_name
BMC Biotechnoljournal_title
BMC biotechnologyauthors
Mukai A,Ichiraku A,Horikawa Kdoi
10.1186/s12896-016-0267-8subject
Has Abstractpub_date
2016-04-14 00:00:00pages
37issn
1472-6750pii
10.1186/s12896-016-0267-8journal_volume
16pub_type
杂志文章abstract:BACKGROUND:Serine hydroxymethyltransferase (SHMT) is the key enzyme in L-serine enzymatic production, suggesting the importance of obtaining a SHMT with high activity. RESULTS:Here, a novel SHMT gene, glyA, was obtained through degenerate oligonucleotide-primed PCR and encoded a novel SHMT with 54.3% similarity to the...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-014-0093-9
更新日期:2014-11-14 00:00:00
abstract:BACKGROUND:Heat-stable antifungal factor (HSAF) is a newly identified broad-spectrum antifungal antibiotic from the biocontrol agent Lysobacter enzymogenes and is regarded as a potential biological pesticide, due to its novel mode of action. However, the production level of HSAF is quite low, and little research has re...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-018-0478-2
更新日期:2018-10-24 00:00:00
abstract:BACKGROUND:Baculovirus, which has a width of 40 nm and a length of 250-300 nm, can display functional peptides, receptors and antigens on its surface by their fusion with a baculovirus envelop protein, GP64. In addition, some transmembrane proteins can be displayed without GP64 fusion, using the native transmembrane do...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-11-60
更新日期:2011-06-02 00:00:00
abstract:BACKGROUND:Interest in using plants for production of recombinant proteins such as monoclonal antibodies is growing, but proteolytic degradation, leading to a loss of functionality and complications in downstream purification, is still a serious problem. RESULTS:In this study, we investigated the dynamics of the assem...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-11-128
更新日期:2011-12-30 00:00:00
abstract:BACKGROUND:There is an increasing need to understand cell-cell interactions for cell and tissue engineering purposes, such as optimizing cell sheet constructs, as well as for examining adhesion defect diseases. For cell-sheet engineering, one major obstacle to sheet function is that cell sheets in suspension are fragil...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-13-17
更新日期:2013-02-26 00:00:00
abstract:BACKGROUND:Copper is an essential chemical element for life as it is a part of prosthetic groups of enzymes including super oxide dismutase and cytochrome c oxidase; however, it is also toxic at high concentrations. Here, we present the trade-off of copper availability and growth inhibition of a common host used for co...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-016-0251-3
更新日期:2016-02-20 00:00:00
abstract:BACKGROUND:The cattle ticks, Boophilus spp., affect cattle production in tropical and subtropical regions of the world. Tick vaccines constitute a cost-effective and environmentally friendly alternative to tick control. The recombinant B. microplus Bm86 protective antigen has been shown to protect cattle against tick i...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-9-29
更新日期:2009-03-31 00:00:00
abstract:BACKGROUND:Whole genome amplification (WGA) promises to eliminate practical molecular genetic analysis limitations associated with genomic DNA (gDNA) quantity. We evaluated the performance of multiple displacement amplification (MDA) WGA using gDNA extracted from lymphoblastoid cell lines (N = 27) with a range of start...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-5-24
更新日期:2005-09-16 00:00:00
abstract:BACKGROUND:The proteomics literature has seen a proliferation of publications that seek to apply the rapidly improving technology of 2D gels to study various biological systems. However, there is a dearth of systematic studies that have investigated appropriate statistical approaches to analyse the data from these expe...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-5-7
更新日期:2005-02-11 00:00:00
abstract:BACKGROUND:Spinal cord injury (SCI) remains a huge medical problem nowadays as there is no hospital providing the versatile strategies for repairing central nervous system and restoring function. Herein, we focused on PC-12 cells as an important research tool and studied the potential role of resveratrol (RSV) in infla...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-019-0502-1
更新日期:2019-01-28 00:00:00
abstract:BACKGROUND:Circular forms of viral genomic DNA are generated during infection of cells with retroviruses like HIV-1. Such circles are unable to replicate and are eventually lost as a result of cell division, lending support to the prevalent notion that episomal retroviral DNA forms are dead-end products of reverse tran...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-8-60
更新日期:2008-08-09 00:00:00
abstract:BACKGROUND:The antiphospholipid syndrome (APS), characterised by recurrent miscarriage and thrombosis, is a significant cause of morbidity and mortality. Domain I (DI) of human beta 2 glycoprotein I (beta2GPI) is thought to contain crucial antibody binding epitopes for antiphospholipid antibodies (aPL), which are criti...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-6-8
更新日期:2006-02-10 00:00:00
abstract:BACKGROUND:Quantum dots (QDs) have been considered as a new and efficient probe for labeling cells non-invasively in vitro and in vivo, but fairly little is known about how QDs are eliminated from cells after labeling. The purpose of this study is to investigate the metabolism of QDs in different type of cells. RESULT...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-10-36
更新日期:2010-05-06 00:00:00
abstract:BACKGROUND:As with many plant species, current genome editing strategies in soybean are initiated by stably transforming a gene that encodes an engineered nuclease into the genome. Expression of the transgene results in a double-stranded break and repair at the targeted locus, oftentimes resulting in mutation(s) at the...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-020-00604-3
更新日期:2020-02-24 00:00:00
abstract:BACKGROUND:Bacterial degradation/transformation of steroids is widely investigated to create biotechnologically relevant strains for industrial application. The strain of Nocardioides simplex VKM Ac-2033D is well known mainly for its superior 3-ketosteroid Δ1-dehydrogenase activity towards various 3-oxosteroids and oth...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-021-00668-9
更新日期:2021-01-13 00:00:00
abstract:BACKGROUND:Mesenchymal stem cells (MSCs) are of great interest in bone regenerative medicine due to their osteogenic potential and trophic effects. However, challenges to large-scale production of MSCs can hinder the translation of MSC therapies. 3D Microcarrier (MC)-based MSC culture presents a scalable and cost-effec...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-015-0219-8
更新日期:2015-10-31 00:00:00
abstract:BACKGROUND:Oleaginous yeast species are an alternative for the production of lipids or triacylglycerides (TAGs). These yeasts are usually non-pathogenic and able to store TAGs ranging from 20 % to 70 % of their cell mass depending on culture conditions. TAGs originating from oleaginous yeasts can be used as the so-call...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-016-0276-7
更新日期:2016-05-27 00:00:00
abstract:BACKGROUND:In the previous study, the cellulolytic Escherichia coli ZH-4 isolated from bovine rumen was found to show extracellular cellulase activity and could degrade cellulose in the culture. The goal of this work was to identify and characterize the secreted cellulase of E. coli ZH-4. It will be helpful to re-under...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-019-0556-0
更新日期:2019-08-27 00:00:00
abstract:BACKGROUND:The ability to produce the same recombinant protein in both prokaryotic and eukaryotic cells offers many experimental opportunities. However, the cloning of the same gene into multiple plasmids is required, which is time consuming, laborious and still may not produce soluble, stable protein in sufficient qua...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-12-54
更新日期:2012-08-23 00:00:00
abstract:BACKGROUND:Transformed cells of Escherichia coli DH5-alpha with pGFPuv, induced by IPTG (isopropyl-beta-d-thiogalactopyranoside), express the green fluorescent protein (gfpuv) during growth phases. E. coli subjected to the combination of selective permeation by freezing/thawing/sonication cycles followed by the three-p...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-2-7
更新日期:2002-04-24 00:00:00
abstract:BACKGROUND:Multi-copy gene integration into microbial genomes is a conventional tool for obtaining improved gene expression. For Penicillium chrysogenum, the fungal producer of the beta-lactam antibiotic penicillin, many production strains carry multiple copies of the penicillin biosynthesis gene cluster. This discover...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-017-0335-8
更新日期:2017-02-16 00:00:00
abstract:BACKGROUND:Tetanus neurotoxin (TeNT) is taken up at nerve terminals and undergoes retrograde migration. The toxic properties of TeNT reside in the toxin light chain (L), but like complete TeNT, the TeNT heavy chain (TTH) and the C-terminal domain (TTC) alone can bind and enter into neurons. Here, we explored whether at...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-018-0452-z
更新日期:2018-06-11 00:00:00
abstract:BACKGROUND:Rapid, robust and reversible induction of transgene expression would significantly facilitate cancer gene therapy as well as allow the in vivo functional study of newly discovered genes in tumor formation and progression. The popularity of the ecdysone inducible gene switch system has led us to investigate w...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-1-11
更新日期:2001-01-01 00:00:00
abstract:BACKGROUND:In spite of a great number of monomeric fluorescent proteins developed in the recent years, the reported fluorescent protein-based FRET pairs are still characterized by a number of disadvantageous features, complicating their use as reporters in cell biology and for high-throughput cell-based screenings. RE...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-9-24
更新日期:2009-03-25 00:00:00
abstract:BACKGROUND:The use of highly reproducible and spatiallyhomogeneous thin film matrices permits automated microscopy and quantitative determination of the response of hundreds of cells in a population. Using thin films of extracellular matrix proteins, we have quantified, on a cell-by-cell basis, phenotypic parameters of...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-6-14
更新日期:2006-03-06 00:00:00
abstract:BACKGROUND:Eleven new human polyomaviruses (HPyVs) have been identified in the last decade. Serological studies show that these novel HPyVs sub-clinically infect humans at an early age. The routes of infection, entry pathways, and cell tropism of new HPyVs remain unknown. VP1 proteins of polyomaviruses can assembly int...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-015-0187-z
更新日期:2015-08-04 00:00:00
abstract:BACKGROUND:The recent H1N1 influenza pandemic illustrated the shortcomings of the vaccine manufacturing process. The A/California/07/2009 H1N1 pandemic influenza vaccine or A(H1N1)pdm09 was available late and in short supply as a result of delays in production caused by low yields and poor antigen stability. Recombinan...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-12-77
更新日期:2012-10-30 00:00:00
abstract:BACKGROUND:The green fluorescent protein (GFP) has been regarded as a valuable tool and widely applied as a biomarker in medical applications and diagnostics. A cost-efficient upstream expression system and an inexpensive downstream purification process will meet the demands of the GFP protein with high-purity. RESULT...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-019-0590-y
更新日期:2019-12-09 00:00:00
abstract:BACKGROUND:Carboxyl-terminal protease (CtpA) plays essential functions in posttranslational protein processing in prokaryotic and eukaryotic cells. To date, only a few bacterial ctpA genes have been characterized. Here we cloned and characterized a novel CtpA. The encoding gene, ctpAp (ctpA of Paenibacillus lautus), wa...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-13-89
更新日期:2013-10-25 00:00:00
abstract:BACKGROUND:Asian rust (Phakopsora pachyrhizi) is a common disease in Brazilian soybean fields and it is difficult to control. To identify a biochemical candidate with potential to combat this disease, a new chitinase-like xylanase inhibitor protein (XIP) from coffee (Coffea arabica) (CaclXIP) leaves was cloned into the...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-11-14
更新日期:2011-02-07 00:00:00