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 adsorption of nucleic acids, especially single-stranded or single-stranded-region-containing nucleic acids, leading to approximately 50% loss of phosphopeptides during the process of IMAC enrichment. Therefore, nucleic acids must be removed from protein samples prior to IMAC. Acetonitrile (ACN) precipitation, a simple and efficient procedure, was established to remove nucleic acids from the protein samples. We showed that ACN precipitation approximately doubled the phosphopeptide number identified by IMAC and mass spectrometry, indicating that nucleic acid removal significantly improves the identification of phosphopeptides.
journal_name
Mol Biotechnoljournal_title
Molecular biotechnologyauthors
Li Y,Luo Y,Wu S,Gao Y,Liu Y,Zheng Ddoi
10.1007/s12033-009-9176-6subject
Has Abstractpub_date
2009-09-01 00:00:00pages
59-66issue
1eissn
1073-6085issn
1559-0305journal_volume
43pub_type
杂志文章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：:Clinical molecular genetics has recently become recognized as a diagnostic discipline. This article covers the evolution, structure, and possible forward development of clinical molecular genetics. Topics covered include general test categories, introducing new tests, laboratory facilities, staffing and training, and ...
journal_title：Molecular biotechnology
pub_type： 杂志文章,评审
doi：10.1007/BF02752254
更新日期：1997-10-01 00:00:00
abstract：:In this review I briefly describe the technique of protein engineering and indicate how the present state of knowledge allows proteins to be mutated to increase or decrease stability. I discuss experiments on both model proteins and those of relevance to the food industry and show how hydrophobic forces are a major dr...
journal_title：Molecular biotechnology
pub_type： 杂志文章,评审
doi：10.1007/BF02921609
更新日期：1995-10-01 00:00:00
abstract：:In this study, we investigated expression and dimerization of an ER-associated degradation (ERAD) substrate, a null Hong Kong variant of α-1-antitrypsin (NHK) using immunoblotting assay and a novel NanoLuc complementary reporter system called the NanoBiT (NB) assay. This NB-tagged NHK made it possible to monitor the i...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-018-0092-5
更新日期：2018-08-01 00:00:00
abstract：:Recombinant proteins are useful tools in biological research, drug development, and drug screening. Specially designed expression vectors have been developed to introduce cDNA for recombinant protein expression in mammalian cells. We have combined a dicistronic mRNA design for expression of the recombinant protein, us...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/BF02745860
更新日期：1998-08-01 00:00:00
abstract：:The monomeric bacteriophage RNA polymerases allow the synthesis of virtually any RNA molecule in unlimited quantity. In this protocol, we describe the preparation of plasmid and PCR-derived templates. A basic transcription protocol is provided with several optional modifications. The use of RNA probes in Northern blot...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/BF02761751
更新日期：1997-04-01 00:00:00
abstract：:Hypermethylated in Cancer-1 (HIC-1) is a novel tumor suppressor plays crucial role in tumor formation through loss of function by hypermethylation. HIC-1 is known as transcriptional factor whereas little known about its structure and function. Requirement felt to clone and express full coding protein and reveal variou...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-014-9783-8
更新日期：2014-11-01 00:00:00
abstract：:HOX genes are highly conserved transcription factors that determine the identity of cells and tissues along the anterior-posterior body axis in developing embryos. Aberrations in HOX gene expression have been shown in various tumors. However, the correlation of HOX gene expression patterns with tumorigenesis and cance...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-013-9682-4
更新日期：2014-01-01 00:00:00
abstract：:Nitrilases have attracted tremendous attention for the preparation of optically pure carboxylic acids. This article aims to address the production and utilization of a highly enantioselective nitrilase from Pseudomonas putida MTCC 5110 for the hydrolysis of racemic mandelonitrile to (R)-mandelic acid. The nitrilase ge...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-008-9094-z
更新日期：2009-01-01 00:00:00
abstract：:Lactic acid bacteria (LAB) are generally sensitive to hydrogen peroxide (H(2)O(2)), Lactobacillus sakei YSI8 is one of the very few LAB strains able to degrade H(2)O(2) through the action of a heme-dependent catalase. Lactobacillus rhamnosus strains are very important probiotic starter cultures in meat product ferment...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-010-9254-9
更新日期：2010-06-01 00:00:00
abstract：:The biosensor field has grown enormously since the first demonstration of the biosensor concept by Leland C. Clark, Jr. in 1962. Today's biosensor market is dominated by glucose biosensors, mass-produced enzyme electrodes for the rapid self-diagnosis of blood glucose levels by diabetes sufferers. Here we take a histor...
journal_title：Molecular biotechnology
pub_type： 杂志文章,评审
doi：10.1385/MB:32:3:249
更新日期：2006-03-01 00:00:00
abstract：:High-quality RNA preparations are critical for further applications such as reverse transcriptase-polymerase chain reaction (RT-PCR) transcript amplifications, and elaboration of cDNA and expressed sequence tag libraries. Melanins are phenolic compounds present in many fungi and apparently play key roles in fungi path...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1385/MB:34:1:45
更新日期：2006-09-01 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：: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：:Artificial microRNA (amiRNA) has become a powerful tool for gene silencing in plants. A new method for easy and rapid construction of rice artificial miRNA vector is described. The procedure involved modification of the pCAMBIA1300-UR vector by insertion of a 'vector modification fragment'. This was prepared from the ...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-010-9291-4
更新日期：2010-11-01 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：:DNA beta is an approx 1350 nucleotide, single-stranded DNA molecule which has been shown to be associated with some monopartite geminiviruses of the genus Begomovirus. This component requires the helper begomovirus for replication in the cells of host plants and for insect transmission, possibly by trans-encapsidation...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1385/MB:20:3:315
更新日期：2002-03-01 00:00:00
abstract：:Virus-like particles (VLPs) have vast potential for applications in nanoscience and nanomedicine. These biological nanoparticles may be used for medical imaging, vaccination, or tissue-specific delivery of drugs or other bioactive molecules. VLPs of Human parvovirus B19 (B19 V) can be assembled in vitro from the recom...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-014-9823-4
更新日期：2015-04-01 00:00:00
abstract：:UV-B radiation can damage biomolecules, such as DNA, RNA, and proteins, halting essential cellular processes; this damage is partly due to ROS generation. Plant secondary metabolites may protect against UV-B. Psychotria brachyceras Müll. Arg. (Rubiaceae), a subtropical shrub, produces brachycerine, a monoterpene indol...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-012-9546-3
更新日期：2013-05-01 00:00:00
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...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1385/MB:30:2:103
更新日期：2005-06-01 00:00:00
abstract：:Positional cloning of rare disease genes depends on the availability of highly polymorphic markers near the disease loci. The most abundant class of polymorphic markers in the human genome is CA-repeats. We have developed a strategy for the rapid isolation of highly polymorphic CA-repeats from YAC clones. Total DNA of...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/BF02789104
更新日期：1995-04-01 00:00:00
abstract：:This article reviews some of the issues involved in evaluating vaccines in humans. Vaccine trials are required for licensure and are essential for demonstrating a vaccine's safety and protective efficacy. The formal framework of phase I, II, and III trials is described, with particular emphasis on the choice of hypoth...
journal_title：Molecular biotechnology
pub_type： 杂志文章,评审
doi：10.1385/MB:17:1:43
更新日期：2001-01-01 00:00:00
abstract：:Fibrillation of α-synuclein proceeds through distinct stages, with oligomers combining to form the seed or the nucleus, followed by exponential and saturation phases. Osmolytes are considered to act as protein stabilizers by virtue of their ability to inhibit protein aggregation. Trehalose, a non-reducing disaccharide...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-016-9923-4
更新日期：2016-04-01 00:00:00
abstract：:CD151 is an abundantly expressed eukaryotic transmembrane protein on the cell surface. It is involved in cell adhesion, angiogenesis and signal transduction as well in disease conditions such as cancer and viral infections. However, the molecular mechanism of CD151 activation is poorly understood due to the lack of st...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-019-00212-3
更新日期：2019-12-01 00:00:00
abstract：:Agrobacterium-mediated transformation is the method of choice to engineer desirable genes into plants. Here we describe a protocol for demonstrating T-DNA transfer from Agrobacterium into the economically important graminaceous plant maize. Expression of the T-DNA-located GUS gene was observed with high efficiency on ...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1385/MB:13:2:165
更新日期：1999-12-01 00:00:00
abstract：:A short (43-bp) A/T-rich stretch of DNA located in the intergenic region between the baiA2 and baiF genes from Eubacterium sp. strain VPI 12708 was amplified by polymerase chain reaction (PCR) and inserted in front of the Shine-Dalgarno (SD) sequences of three inefficiently-expressed Eubacterium sp. strain VPI 12708 g...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/BF02789174
更新日期：1999-02-01 00:00:00
abstract：:As the use of adenoviral vectors in gene therapy protocols increases, there is a corresponding need for rapid, accurate, and reproducible titer methods. Multiple methods currently exist for determining titers of recombinant adenoviral vector, including optical absorbance, electron microscopy, fluorescent focus assay, ...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1385/MB:14:3:197
更新日期：2000-03-01 00:00:00
abstract：:Lactococcus lactis is industrially important microorganism used in many dairy fermentations. Numerous genes and gene expression signals from this organism have now been identified and characterized. Recently, several naturally occurring, inducible gene-expression systems have also been described in L. lactis. The main...
journal_title：Molecular biotechnology
pub_type： 杂志文章,评审
doi：10.1007/BF02760814
更新日期：1998-04-01 00:00:00
abstract：:The recombinant acidstable α-amylase (Ba-amy) of acidophilic bacterium Bacillus acidicola TSAS1 has been produced extracellularly using a combination of cloning (E. coli and P. pastoris) and physico-chemical treatment strategies. A total of 150,000 U/L of Ba-amy were attained under constitutive promoter in P. pastoris...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-016-9970-x
更新日期：2016-11-01 00:00:00
abstract：:A naturally secreted Gaussia luciferase (Gluc) has been utilized as a reporter for bioluminescence imaging (BLI) evaluation. However, the potential application of Gluc for in vivo monitoring of systemic protein delivery, as well as its natural biodistribution, has not been studied. To examine Gluc secretion and uptake...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-012-9519-6
更新日期：2013-01-01 00:00:00
