Abstract:
BACKGROUND:The selection of bacterial recombinants that harbour a desired insert, has been a key factor in molecular cloning and a series of screening procedures need to be performed for selection of clones carrying the genes of interest. The conventional cloning techniques are reported to have problems such as screening high number of colonies, generation of false positives, setting up of control ligation mix with vector alone etc. RESULTS:We describe the development of a novel dual cloning/expression vector, which enables to screen the recombinants directly and expression of the gene of interest. The vector contains Green fluorescence protein (GFP) as the reporter gene and is constructed in such a way that the E. coli cells upon transformation with this vector does not show any fluorescence, but readily fluoresce upon insertion of a foreign gene of interest. The same construct could be easily used for screening of the clones and expression studies by mere switching to specific hosts. CONCLUSIONS:This is the first vector reported that takes the property of colour or fluorescence to be achieved only upon cloning while all the other vectors available commercially show loss of colour or loss of fluorescence upon cloning. As the fluorescence of GFP depends on the solubility of the protein, the intensity of the fluorescence would also indicate the extent of solubility of the expressed target protein.
journal_name
Microb Cell Factjournal_title
Microbial cell factoriesauthors
Banerjee S,Kumar J,Apte-Deshpande A,Padmanabhan Sdoi
10.1186/1475-2859-9-30subject
Has Abstractpub_date
2010-05-11 00:00:00pages
30issn
1475-2859pii
1475-2859-9-30journal_volume
9pub_type
杂志文章abstract:BACKGROUND:Fumaric acid is a commercially important component of foodstuffs, pharmaceuticals and industrial materials, yet the current methods of production are unsustainable and ecologically destructive. RESULTS:In this study, the fumarate biosynthetic pathway involving reductive reactions of the tricarboxylic acid c...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-11-24
更新日期:2012-02-15 00:00:00
abstract::All of humans and other mammalian species are colonized by some types of microorganisms such as bacteria, archaea, unicellular eukaryotes like fungi and protozoa, multicellular eukaryotes like helminths, and viruses, which in whole are called microbiota. These microorganisms have multiple different types of interactio...
journal_title:Microbial cell factories
pub_type: 杂志文章,评审
doi:10.1186/s12934-020-01483-1
更新日期:2020-11-26 00:00:00
abstract:: Different species of microorganisms including yeasts, filamentous fungi and bacteria have been used in the past 25 years for the controlled production of foreign proteins of scientific, pharmacological or industrial interest. A major obstacle for protein production processes and a limit to overall success has been th...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-7-11
更新日期:2008-04-04 00:00:00
abstract:BACKGROUND:The classic AOX1 replacement approach is still one of the most often used techniques for expression of recombinant proteins in the methylotrophic yeast Pichia pastoris. Although this approach is largely successful, it frequently delivers clones with unpredicted production characteristics and a work-intense s...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-016-0486-7
更新日期:2016-05-20 00:00:00
abstract:BACKGROUND:The rational design of L-phenylalanine (L-Phe) overproducing microorganisms has been successfully achieved by combining different genetic strategies such as inactivation of the phosphoenolpyruvate: phosphotransferase transport system (PTS) and overexpression of key genes (DAHP synthase, transketolase and cho...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-6-30
更新日期:2007-09-19 00:00:00
abstract:BACKGROUND:L-arabinose is an important intermediate for anti-virus drug synthesis and has also been used in food additives for diets-controlling in recent years. Commercial production of L-arabinose is a complex progress consisting of acid hydrolysis of gum arabic, followed by multiple procedures of purification, thus ...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-10-43
更新日期:2011-06-07 00:00:00
abstract:BACKGROUND:We evaluated the functional capacity of plantaricin-producing Lactobacillus plantarum SF9C and S-layer-carrying Lactobacillus brevis SF9B to withstand gastrointestinal transit and to compete among the gut microbiota in vivo. Considering the probiotic potential of Lb. brevis SF9B, this study aims to investiga...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-020-01365-6
更新日期:2020-05-19 00:00:00
abstract:BACKGROUND:Chinese Nong-flavor (NF) liquor is continuously and stably produced by solid-state fermentation technology for 1000 years, resulting in enrichment of special microbial community and enzymes system in its starter. Based on traditional culture-dependent methods, these functional enzymes are hardly obtained. Ac...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-018-0878-y
更新日期:2018-02-22 00:00:00
abstract:BACKGROUND:The discovery of the autotransporter family has provided a mechanism for surface expression of proteins in laboratory strains of Escherichia coli. We have previously reported the use of the AIDA-I autotransport system to express the Salmonella enterica serovar Enteritidis proteins SefA and H:gm. The SefA pro...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-11-118
更新日期:2012-09-03 00:00:00
abstract:BACKGROUND:Our laboratory has constructed a Bacillus stearothermophilus α-amylase (AmyS) derivative with excellent enzymatic properties. Bacillus subtilis is generally regarded as safe and has excellent protein secretory capability, but heterologous extracellular production level of B. stearothermophilus α-amylase in B...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-019-1119-8
更新日期:2019-04-11 00:00:00
abstract:BACKGROUND:In Saccharomyces cerevisiae, alpha-glucosidase (maltase) is a key enzyme in maltose metabolism. In addition, the overexpression of the alpha-glucosidase-encoding gene MAL62 has been shown to increase the freezing tolerance of yeast in lean dough. However, its cryoprotection mechanism is still not clear. RES...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-020-01454-6
更新日期:2020-10-19 00:00:00
abstract:BACKGROUND:The E. coli lac operon and its components have been studied for decades, and lac-derived systems are widely used for recombinant protein production. However, lac operon dynamics and induction behavior remain the paradigm of gene regulation. Recently, an HPLC-MS-based method to quantify IPTG in the medium and...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-11-58
更新日期:2012-05-09 00:00:00
abstract:BACKGROUND:Pichia pastoris is one of the most important host organisms for the recombinant production of proteins in industrial biotechnology. To date, strain specific parameters, which are needed to set up feeding profiles for fed batch cultivations, are determined by time-consuming continuous cultures or consecutive ...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-10-14
更新日期:2011-03-03 00:00:00
abstract:BACKGROUND:Raspberry ketone is the primary aroma compound found in raspberries and naturally derived raspberry ketone is a valuable flavoring agent. The economic incentives for the production of raspberry ketone, combined with the very poor yields from plant tissue, therefore make this compound an excellent target for ...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-016-0446-2
更新日期:2016-03-04 00:00:00
abstract:BACKGROUND:Targeting of cellular proteins to the extracellular environment is directed by a secretory signal sequence located at the N-terminus of a secretory protein. These signal sequences usually contain an N-terminal basic amino acid followed by a stretch containing hydrophobic residues, although no consensus signa...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-015-0203-y
更新日期:2015-02-14 00:00:00
abstract:BACKGROUND:The production of high yields of recombinant proteins is an enduring bottleneck in the post-genomic sciences that has yet to be addressed in a truly rational manner. Typically eukaryotic protein production experiments have relied on varying expression construct cassettes such as promoters and tags, or cultur...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-8-10
更新日期:2009-01-29 00:00:00
abstract:BACKGROUND:Amylases are used in various industrial processes and a key requirement for the efficiency of these processes is the use of enzymes with high catalytic activity at ambient temperature. Unfortunately, most amylases isolated from bacteria and filamentous fungi have optimal activity above 45 °C and low pH. For ...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-017-0693-x
更新日期:2017-05-02 00:00:00
abstract:BACKGROUND:The budding yeast Pichia pastoris is widely used for protein production. To determine the best suitable strategy for strain improvement, especially for high secretion, quantitative data of intracellular fluxes of recombinant protein are very important. Especially the balance between intracellular protein for...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-10-47
更新日期:2011-06-26 00:00:00
abstract:BACKGROUND:L-tryptophan is an aromatic amino acid widely used in the food, chemical and pharmaceutical industries. In Escherichia coli, L-tryptophan is synthesized from phosphoenolpyruvate and erythrose 4-phosphate by enzymes in the shikimate pathway and L-tryptophan branch pathway, while L-serine and phosphoribosylpyr...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-11-30
更新日期:2012-03-02 00:00:00
abstract:BACKGROUND:The proteome reflects the available cellular machinery to deal with nutrients and environmental challenges. The most common E. coli strain BL21 growing in different, commonly employed media was evaluated using a detailed quantitative proteome analysis. RESULTS:The presence of preformed biomass precursor mol...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-13-45
更新日期:2014-03-23 00:00:00
abstract:BACKGROUND:Catalase (EC 1.11.1.6) is one of the important industrial enzyme employed in diagnostic and analytical methods in the form of biomarkers and biosensors in addition to their enormous applications in textile, paper, food and pharmaceutical sectors. The present study demonstrates the utility of a newly isolated...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-016-0410-1
更新日期:2016-01-14 00:00:00
abstract::L-arginine (ARG) is an important amino acid for both medicinal and industrial applications. For almost six decades, the research has been going on for its improved industrial level production using different microorganisms. While the initial approaches involved random mutagenesis for increased tolerance to ARG and con...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-014-0166-4
更新日期:2014-12-03 00:00:00
abstract:BACKGROUND:The filamentous fungus T. reesei effectively degrades cellulose and is known to produce various cellulolytic enzymes such as beta-glucosidase, endoglucanase, and cellobiohydrolase. The expression levels of each cellulase are controlled simultaneously, and their ratios and synergetic effects are important for...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-9-32
更新日期:2010-05-14 00:00:00
abstract:BACKGROUND:The methylotrophic yeast Pichia pastoris is a well-studied host organism for recombinant protein production, which is usually regulated either by a constitutive promoter (e.g. promoter of glyceraldehyde-3-phosphate dehydrogenase; PGAP) or an inducible promoter (e.g. promoter of alcohol oxidase 1; PAOX1). Bot...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-017-0768-8
更新日期:2017-09-13 00:00:00
abstract::Lactococcus lactis has progressed a long way since its discovery and initial use in dairy product fermentation, to its present biotechnological applications in genetic engineering for the production of various recombinant proteins and metabolites that transcends the heterologous species barrier. Key desirable features...
journal_title:Microbial cell factories
pub_type: 杂志文章,评审
doi:10.1186/s12934-017-0669-x
更新日期:2017-04-04 00:00:00
abstract:BACKGROUND:A key advantage of recombinant antibody technology is the ability to optimize and tailor reagents. Single domain antibodies (sdAbs), the recombinantly produced variable domains derived from camelid and shark heavy chain antibodies, provide advantages of stability and solubility and can be further engineered ...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-017-0837-z
更新日期:2017-12-12 00:00:00
abstract:BACKGROUND:Escherichia coli has emerged as a promising platform microorganism to produce biofuels and fine chemicals of industrial interests. Certain obstacles however remain to be overcome, among which organic-solvent tolerance is a crucial one. RESULTS:We used global transcription machinery engineering (gTME) to imp...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-015-0368-4
更新日期:2015-11-05 00:00:00
abstract:BACKGROUND:In spite of the substantial metabolic engineering effort previously devoted to the development of Saccharomyces cerevisiae strains capable of fermenting both the hexose and pentose sugars present in lignocellulose hydrolysates, the productivity of reported strains for conversion of the naturally most abundan...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-9-16
更新日期:2010-03-10 00:00:00
abstract:BACKGROUND:The expression of heterologous proteins in Escherichia coli is strongly affected by codon bias. This phenomenon occurs when the codon usage of the mRNA coding for the foreign protein differs from that of the bacterium. The ribosome pauses upon encountering a rare codon and may detach from the mRNA, thereby t...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-8-41
更新日期:2009-07-24 00:00:00
abstract:BACKGROUND:The oleaginous yeast Yarrowia lipolytica is an organism of choice for the tailored production of various compounds such as biofuels or biopolymers. When properly engineered, it is capable of producing medium-chain-length polyhydroxyalkanoate (mcl-PHA), a biobased and biodegradable polymer that can be used as...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-019-1140-y
更新日期:2019-05-31 00:00:00