Abstract：
:Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a hematopoietic growth factor, that has been used as a therapeutic agent in facilitating bone marrow and stem cell transplantation and in other clinical cases like neutropenia. Although biologically active recombinant GM-CSF has been successfully produced in Escherichia coli, the reported levels are extremely poor. In this study we looked into the possible reasons for poor expression and found that protein toxicity coupled with protease-based degradation was the principal reason for low productivity. To overcome this problem we attached a signal sequence, as well as an amino-terminal His-tag fusion to the GM-CSF gene. This combination had a dramatic effect on expression levels, which increased from 0.8 microg/mL in the control to 40 microg/mL. When a larger fusion partner, such as the Maltose-binding protein (MBP-tag), was used the expression levels increased further to 69.5 microg/mL, which along with the MBP-tag represented approx 12% of the total cellular protein.
journal_name
Mol Biotechnoljournal_title
Molecular biotechnologyauthors
Bhattacharya P,Pandey G,Srivastava P,Mukherjee KJdoi
10.1385/MB:30:2:103keywords:
subject
Has Abstractpub_date
2005-06-01 00:00:00pages
103-16issue
2eissn
1073-6085issn
1559-0305pii
MB:30:2:103journal_volume
30pub_type
杂志文章abstract：:The clinical management of immunocompromised patients depends on the rapid identification of infectious agents such as fungal pathogens. The procedure described here for accomplishing this uses a sensitive polymerase chain reaction method, previously reported, combined with restriction-enzyme digestion to distinguish ...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1385/MB:24:3:229
更新日期：2003-07-01 00:00:00
abstract：:Deficiency of the lysosomal glucocerebrosidase (GCR) enzyme results in Gaucher's disease, the most common inherited storage disorder. Treatment consists of enzyme replacement therapy by the administration of recombinant GCR produced in Chinese hamster ovary cells. The production of anti-GCR antibodies has already been...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-010-9303-4
更新日期：2010-11-01 00:00:00
abstract：:The use of synthetic genes may constitute a successful approach for the heterologous production and functional expression of bacterial antimicrobial peptides (bacteriocins) by recombinant yeasts. In this work, synthetic genes with adapted codon usage designed from the mature amino acid sequence of the bacteriocin ente...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-014-9731-7
更新日期：2014-06-01 00:00:00
abstract：:Coptotermes curvignathus is a termite that, owing to its ability to digest living trees, serves as a gold mine for robust industrial enzymes. This unique characteristic reflects the presence of very efficient hydrolytic enzyme systems including cellulases. Transcriptomic analyses of the gut of C. curvignathus revealed...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-017-0015-x
更新日期：2017-07-01 00:00:00
abstract：:In mammals, CpG methylation is one of the mechanisms of epigenetic control over the linear sequence of bases of deoxyribonucleic acid (DNA); about 70% of CpG dinucleotides are methylated. The actual signal that triggers DNA methylation is not known, although repetitive DNA has been shown to be an attractive template f...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1385/MB:26:3:215
更新日期：2004-03-01 00:00:00
abstract：:The diversity and complexity of bioinformatics tools currently available for protein sequence analysis can make it difficult to know where to begin when presented with a new sequence. In this article, we present a protocol outlining one approach to sequence analysis that should give as comprehensive a picture as possi...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-007-0032-2
更新日期：2007-07-01 00:00:00
abstract：:The electron microscopic study of DNA-protein complexes can yield valuable information that is often not easily available by other methods. In this article we give a number of examples that were chosen to illustrate the utility of electron microscopy. Along with the strategy used are protocols that allow such experime...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1385/MB:16:1:77
更新日期：2000-09-01 00:00:00
abstract：:The macrofungus Cordyceps militaris contains many kinds of bioactive ingredients that are regulated by functional genes, but the functions of many genes in C. militaris are still unknown. In this study, to improve the frequency of homologous integration, a genetic transformation system based on a split-marker approach...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-018-0080-9
更新日期：2018-05-01 00:00:00
abstract：:To develop combined vaccine systems, we have generated Edwardsiella tarda ghosts (ETG) displaying a foreign protein on the outer membrane and also Ed. tarda cadaver (ETC) expressing a heterologous protein in the cytoplasm. Green fluorescent protein (GFP) was used as a model foreign protein. A constitutive promoter (Et...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-010-9277-2
更新日期：2010-10-01 00:00:00
abstract：:Due to the limited number of molecular studies focused on European gene pool investigation, it is necessary to perform plant material recognition. Eighteen accessions of three Miscanthus species, namely, M. × giganteus, M. sinensis, M. sacchariflorus were evaluated with the use of molecular marker systems such as: int...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-014-9770-0
更新日期：2014-10-01 00:00:00
abstract：:Immobilized-metal-ion affinity chromatography (IMAC) is used extensively for phosphopeptide enrichment in phosphoproteomics. However, the effect of nucleic acids in protein samples on phosphopeptide enrichment by IMAC has not yet been well clarified. In this study, we demonstrate that IMAC beads possess a strong adsor...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-009-9176-6
更新日期：2009-09-01 00:00:00
abstract：:De novo design provides an in silico toolkit for the design of novel small molecular structures to a set of specified structural constraints. With the avalanche of bioinformatics data, de novo design is ideally suited for exploring molecules that could be useful for chemical genomics. The design process involves manip...
journal_title：Molecular biotechnology
pub_type： 杂志文章,评审
doi：10.1007/s12033-007-0037-x
更新日期：2007-11-01 00:00:00
abstract：:In order to further study the B subunit of the Escherichia coli heat-labile enterotoxin (LTB), we obtained the LTB gene from pathogenic E. coli, cloned it into the pET22b (+) prokaryotic expression vector, and expressed it as a fusion protein with His tag in E. coli BL21 (DE3). The recombinant LTB was expressed and pu...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-008-9101-4
更新日期：2009-02-01 00:00:00
abstract：:The simple use of nonisotopic hybridization probes to detect complementary sequences provides valuable information in a large number of research and commercial applications. In hybridization assays, the four "S's (speed, simplicity, sensitivity, and specificity) are important criteria for determining the choice of pro...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/BF02740771
更新日期：1996-10-01 00:00:00
abstract：:Adeno-associated virus (AAV) is a small, non-enveloped virus used as vector in gene therapy, mainly produced in human cells and in baculovirus systems. Intense studies on these platforms led to the production of vectors with titers between 103 and 105 viral genomes (vg) per cells. In spite of this, vector yields need ...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-020-00294-4
更新日期：2021-02-01 00:00:00
abstract：:Malaria is a serious but preventable and treatable infectious disease that is found in over 100 countries around the world. Correct and rapid diagnosis of malaria infection can rescue the patient of getting sicker and reduces the risk of disease spreading among humans. Chlamydomonas reinhardtii chloroplast is an attra...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-018-0140-1
更新日期：2019-02-01 00:00:00
abstract：:Addition of taxol into suspension cultures of Taxus cuspidata induced cell apoptosis, which was confirmed by gel electrophoresis of the DNA ladders indicating the progressive delineation of fragmented nuclear DNA (nDNA) into distinct bodies. The additional taxol not only changed the microtubule assembly of cells, but ...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1385/MB:20:2:137
更新日期：2002-02-01 00:00:00
abstract：:A duplex real-time PCR assay with melting curve analysis, using the EvaGreen fluorescence dye, was developed for rapid and reliable identification of bovine and caprine in ruminant feeds. The method merges the use of bovine (Bos taurus) and caprine (Capra hircus) specific primers that amplify small fragments (bovine 9...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-014-9756-y
更新日期：2014-08-01 00:00:00
abstract：:Lentivirus-mediated RNA interference (RNAi) is a potent experimental tool for investigating gene functions in vitro and in vivo. It has advantages that transgenic technology lacks. However, in vivo applications are difficult to apply in the central nervous system of non-model organisms due to the lack of a standard br...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-012-9596-6
更新日期：2013-06-01 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：:Low molecular weight RNA (LMW RNA) is generally obtained either from the total RNA or from total nucleic acids solution. Many steps and chemical reagents are involved in traditional methods for LMW RNA isolation where degradation of LMW RNA often occurs, especially for plant materials with high levels of secondary cat...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-009-9204-6
更新日期：2010-01-01 00:00:00
abstract：:L-Cysteine desulfhydrase (DES; EC 4.4.1.1) is the most important enzyme that catalyzes the decomposition of L-cysteine to pyruvate, ammonia, and hydrogen sulfide (H2S), the latter of which has recently been recognized as the third gasotransmitter for multiple signaling events in plants. Previous results showed the exi...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-012-9621-9
更新日期：2013-07-01 00:00:00
abstract：:The significance of actin cytoskeleton on cell growth was historically studied using toxic drugs, such as cytochalasin. However, it is possible that unpredictable effects of these agents may have influenced the reported observations. In our study, we have established a drug-free system using cofilin overexpression to ...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1385/MB:31:1:001
更新日期：2005-09-01 00:00:00
abstract：:The GacS/A system in Azotobacter vinelandii regulates alginate and alkylresorcinols production through RsmZ1, a small regulatory RNA (sRNA) that releases the translational repression of the algD and arpR mRNAs caused by the RsmA protein. In the Pseudomonadaceae family, the Rsm-sRNAs are grouped into three families: Rs...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-018-0102-7
更新日期：2018-09-01 00:00:00
abstract：:Classic toxicology studies often utilize in vivo animal models. Newer approaches employing in vitro organ-specific cellular models have been developed in recent years to help accelerate the speed and reduce the cost of traditional toxicology testing. Toward the goal of supporting in vitro cellular model research with ...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-020-00245-z
更新日期：2020-04-01 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：:In the context of clinical gene transfer using viral vectors, the risk of memory antivector immunity is often poorly appreciated. The immunological past of the patient, the site of injection, and the vector dose will play intertwined and decisive roles in the safety and efficacy of treatment. To circumvent the drawbac...
journal_title：Molecular biotechnology
pub_type： 杂志文章,评审
doi：10.1385/MB:34:2:247
更新日期：2006-10-01 00:00:00
abstract：:The two asparagine-linked glycosylation sites of recombinant coagulation factor VIIa have been characterized by glycosidase digestions, size-exclusion chromatography (SEC), and mass spectrometry (MS). Nine structures were characterized as core fucosylated bi-and triantennary structures with 0-3 sialic-acid residues wh...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：
更新日期：1998-06-01 00:00:00
abstract：:This review outlines the various methods available for submitting sequence data to the EMBL Nucleotide Sequence Database. Depending on the type of sequence data and the facilities available to the submitter, one method may be more suitable than another. Recent developments have been the World Wide Web submission tool,...
journal_title：Molecular biotechnology
pub_type： 杂志文章,评审
doi：10.1007/BF02762322
更新日期：1996-08-01 00:00:00
abstract：:Ribosome display is a powerful engineering research tool for the high-throughput selection of peptides or proteins, which results in the generation of high-performance binders against nearly any antigen of interest. As a cell-free display system, ribosome display has been well developed with many outstanding achieveme...
journal_title：Molecular biotechnology
pub_type： 杂志文章,评审
doi：10.1007/s12033-018-0133-0
更新日期：2019-01-01 00:00:00
