Abstract:
BACKGROUND:The insect cell line is a critical component in the production of recombinant proteins in the baculovirus expression system and new cell lines hold the promise of increasing both quantity and quality of protein production. RESULTS:Seventy cell lines were established by single-cell cloning from a primary culture of cells derived from eggs of the black witch moth (Ascalapha odorata; Lepidoptera, Noctuidae). Among 8 rapidly growing lines, cell line 38 (Ao38) was selected for further analysis, based on susceptibility to AcMNPV infection and production of secreted alkaline phosphatase (SEAP) from a baculovirus expression vector. In comparisons with low-passage High Five (BTI-Tn-5B1-4) cells, infected Ao38 cells produced beta-galactosidase and SEAP at levels higher (153% and 150%, respectively) than those measured from High Five cells. Analysis of N-glycans of SEAP produced in Ao38 cells revealed two N-glycosylation sites and glycosylation patterns similar to those reported for High Five and Sf9 cells. Glycopeptide isoforms consisted of pauci- or oligomannose, with and without fucose on N-acetylglucosamine(s) linked to asparagine residues. Estimates of Ao38 cell volume suggest that Ao38 cells are approximately 2.5x larger than Sf9 cells but only approximately 74% of the size of High Five cells. Ao38 cells were highly susceptible to AcMNPV infection, similar to infectivity of Sf9 cells. Production of infectious AcMNPV budded virions from Ao38 cells peaked at approximately 4.5 x 10(7) IU/ml, exceeding that from High Five cells while lower than that from Sf9 cells. Ao38 cells grew rapidly in stationary culture with a population doubling time of 20.2 hr, and Ao38 cells were readily adapted to serum-free medium (Sf-900III) and to a suspension culture system. Analysis of Ao38 and a parental Ascalapha odorata cell line indicated that these lines were free of the alphanodavirus that was recently identified as an adventitious agent in High Five cell lines. CONCLUSIONS:Ao38 cells represent a highly productive new insect cell line that will be useful for heterologous protein expression and other applications in biotechnology.
journal_name
BMC Biotechnoljournal_title
BMC biotechnologyauthors
Hashimoto Y,Zhang S,Blissard GWdoi
10.1186/1472-6750-10-50subject
Has Abstractpub_date
2010-07-05 00:00:00pages
50issn
1472-6750pii
1472-6750-10-50journal_volume
10pub_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: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:Xylanase can replace chemical additives to improve the volume and sensory properties of bread in the baking. Suitable baking xylanase with improved yield will promote the application of xylanase in baking industry. The xylanase XYNZG from the Plectosphaerella cucumerina has been previously characterized by h...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-014-0107-7
更新日期:2014-12-16 00:00:00
abstract:BACKGROUND:The ubiquitin protein is present in all eukaryotic cells and promoters from ubiquitin genes are good candidates to regulate the constitutive expression of transgenes in plants. Therefore, two switchgrass (Panicum virgatum L.) ubiquitin genes (PvUbi1 and PvUbi2) were cloned and characterized. Reporter constru...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-11-74
更新日期:2011-07-11 00:00:00
abstract:BACKGROUND:L-ornithine is effective in the treatment of liver diseases and helps strengthen the heart. The commercial applications mean that efficient biotechnological production of L-ornithine has become increasingly necessary. Adaptive evolution strategies have been proven a feasible and efficient technique to achiev...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-13-47
更新日期:2013-06-01 00:00:00
abstract:BACKGROUND:There is an imperative necessity for alternative sources of energy able to reduce the world dependence of fossil oil. One of the most successful options is ethanol obtained mainly from sugarcane and corn fermentation. The foremost residue from sugarcane industry is the bagasse, a rich lignocellulosic raw mat...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-13-94
更新日期:2013-10-31 00:00:00
abstract:BACKGROUND:Porcine reproductive and respiratory syndrome virus (PRRSV) causes major economic losses in the pig industry worldwide. In vivo, the virus infects a subpopulation of tissue macrophages. In vitro, PRRSV only replicates in primary pig macrophages and African green monkey kidney derived cells, such as Marc-145....
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-10-48
更新日期:2010-06-29 00:00:00
abstract:BACKGROUND:Genetically encoded sensors developed on the basis of green fluorescent protein (GFP)-like proteins are becoming more and more popular instruments for monitoring cellular analytes and enzyme activities in living cells and transgenic organisms. In particular, a number of Ca2+ sensors have been developed, eith...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-7-37
更新日期:2007-06-29 00:00:00
abstract:BACKGROUND:Alkaline phosphatase (AP) catalyzes the hydrolytic cleavage of phosphate monoesters under alkaline conditions and plays important roles in microbial ecology and molecular biology applications. Here, we report on the first isolation and biochemical characterization of a thermolabile AP from a metagenome. RES...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-015-0115-2
更新日期:2015-01-31 00:00:00
abstract:BACKGROUND:In a previous study, anthocyanin levels in potato plants were increased by manipulating genes connected with the flavonoid biosynthesis pathway. However, starch content and tuber yield were dramatically reduced in the transgenic plants, which over-expressed dihydroflavonol reductase (DFR). RESULTS:Transgeni...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-8-25
更新日期:2008-03-05 00:00:00
abstract:BACKGROUND:The presence of coliform bacteria is routinely assessed to establish the microbiological safety of water supplies and raw or processed foods. Coliforms are a group of lactose-fermenting Enterobacteriaceae, which most likely acquired the lacZ gene by horizontal transfer and therefore constitute a polyphyletic...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-015-0168-2
更新日期:2015-06-04 00:00:00
abstract:BACKGROUND:von Willebrand factor (VWF) is a key load bearing domain for mamalian cell adhesion by binding various macromolecular ligands in extracellular matrix such as, collagens, elastin, and glycosaminoglycans. Interestingly, vWF like domains are also commonly found in load bearing systems of marine organisms such a...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-016-0247-z
更新日期:2016-02-16 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:Advances in optical imaging modalities and the continued evolution of genetically-encoded fluorescent proteins are coming together to facilitate the study of cell behavior at high resolution in living organisms. As a result, imaging using autofluorescent protein reporters is gaining popularity in mouse trans...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-4-33
更新日期:2004-12-24 00:00:00
abstract:BACKGROUND:Microarrays are powerful tools for DNA-based molecular diagnostics and identification of pathogens. Most target a limited range of organisms and are based on only one or a very few genes for specific identification. Such microarrays are limited to organisms for which specific probes are available, and often ...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-10-13
更新日期:2010-02-17 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:With the increasing number of GMOs on the global market the maintenance of European GMO regulations is becoming more complex. For the analysis of a single food or feed sample it is necessary to assess the sample for the presence of many GMO-targets simultaneously at a sensitive level. Several methods have be...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-12-4
更新日期:2012-01-19 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:Methylated CpG dinucleotides in promoters are associated with the loss of gene expression in recombinant Chinese hamster ovary (CHO) cells during large-scale commercial manufacturing. We evaluated a promoter devoid of CpG dinucleotides, CpGfree, in parallel with a similar CpG containing promoter, CpGrich, fo...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-016-0300-y
更新日期:2016-10-18 00:00:00
abstract:BACKGROUND:At first glance, there appears to be a high degree of similarity between the metabolism of yeast (the Crabtree effect) and human cancer cells (the Warburg effect). At the root of both effects is accelerated metabolic flow through glycolysis which leads to overflows of ethanol and lactic acid, respectively. I...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-017-0362-5
更新日期:2017-05-08 00:00:00
abstract:BACKGROUND:Loop mediated isothermal amplification of nucleic acid templates is a rapid, sensitive and specific method suitable for molecular diagnostics. However the complexity of primer design and the number of primers involved can lead to false positives from non-specific primer interactions. Standard methods of LAMP...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-019-0549-z
更新日期:2019-08-01 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:A modification of a standard method of fluorescence in situ hybridisation (FISH) is described, by which a combination of several substrates and probes on single microscope slides enables more accurate comparisons of the distribution and abundance of chromosomal sequences and improves the relatively low throu...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-6-20
更新日期:2006-03-20 00:00:00
abstract:BACKGROUND:Expression vector engineering technology is one of the most convenient and timely method for cell line development to meet the rising demand of novel production cell line with high productivity. Destabilization of dihydrofolate reductase (dhfr) selection marker by addition of AU-rich elements and murine orni...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-015-0145-9
更新日期:2015-06-02 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: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:One of the most commonly used vectors for gene therapy is the adenoviral vector; its ability to tightly regulate transgene expression is critical for optimizing therapeutic outcomes. The tetracycline-regulated system (especially the Tet-On system) for gene expression is one of the most valuable tools for con...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-015-0121-4
更新日期:2015-02-13 00:00:00
abstract:BACKGROUND:The CRISPR-Cas9 system is a powerful and versatile tool for crop genome editing. However, achieving highly efficient and specific editing in polyploid species can be a challenge. The efficiency and specificity of the CRISPR-Cas9 system depends critically on the gRNA used. Here, we assessed the activities and...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-019-0565-z
更新日期:2019-10-30 00:00:00
abstract:BACKGROUND:Wax esters are important ingredients in cosmetics, pharmaceuticals, lubricants and other chemical industries due to their excellent wetting property. Since the naturally occurring wax esters are expensive and scarce, these esters can be produced by enzymatic alcoholysis of vegetable oils. In an enzymatic rea...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-7-53
更新日期:2007-08-30 00:00:00
abstract:BACKGROUND:Heparan sulfate glycosaminoglycans are diverse components of certain proteoglycans and are known to interact with growth factors as a co-receptor necessary to induce signalling and growth factor activity. In this report we characterize heterogeneously glycosylated recombinant human perlecan domain 1 (HSPG2 a...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-10-66
更新日期:2010-09-09 00:00:00