Abstract：
:Over the last decade transgenic mouse models have become a common experimental tool for unraveling gene function. During this time there has been a growing expectation that transgenes resemble the in vivo state as much as possible. To this end, a preference away from heterologous promoters has emerged, and transgene constructs often utilize the endogenous promoter and gene sequences in BAC, PAC and YAC form without the addition of selectable markers, or at least their subsequent removal. There has been a trend toward controlled integration by homologous recombination, either at a characterized chromosomal localization or in some cases within the allele of interest. Markers such as green fluorescent protein (GFP), beta-galactosidase (LacZ), and alkaline phosphatase (AP) continue to be useful to trace transgenic cells, or transgene expression. The development of technologies such as RNA interference (RNAi), are introducing new ways of using transgenic models. Future developments in RNAi technology may revolutionize tissue specific inactivation of gene function, without the requirement of generating conditionally targeted mice and tissue specific recombinase mice. Transgenic models are biological tools that aid discovery. Overall, the main consideration in the generation of transgenic models is that they are bona fide biological models that best impart the disease model or biological function of the gene that they represent. The main consideration is to make the best model for the biological question at heart and this review aims to simplify that task somewhat. Here we take a historical perspective on the development of transgenic models, with many of the important considerations to be made in design and development along the way.
journal_name
Mol Biotechnoljournal_title
Molecular biotechnologyauthors
Ristevski Sdoi
10.1385/MB:29:2:153keywords:
subject
Has Abstractpub_date
2005-02-01 00:00:00pages
153-63issue
2eissn
1073-6085issn
1559-0305pii
MB:29:2:153journal_volume
29pub_type
杂志文章,评审abstract：:DNA damage is one of the most impactful events in living organisms, leading to DNA sequence changes (mutation) and disruption of biological processes. A study has identified a protein called Damage Suppressor Protein (Dsup) in the tardigrade Ramazzotius varieornatus that has shown to reduce the effects of radiation da...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-020-00273-9
更新日期：2020-12-01 00:00:00
abstract：:Derivation of cardiomyocytes directly from patients' own fibroblasts could offer a new therapeutic approach for those with ischemic heart disease. An essential step toward clinical application is to establish safe conversion of human fibroblasts into a cardiac fate. Here we aimed to efficiently and safely generate car...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-017-0007-x
更新日期：2017-06-01 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：:Cardiomyocytes are terminally differentiated cells that to date have been characterized as poor targets for nonviral gene transfer. This study was therefore designed to determine the optimal nonviral gene transfer parameters in cell cultures of neonatal rat cardiomyocytes and to compare them with the efficiency of gen...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1385/MB:28:1:21
更新日期：2004-09-01 00:00:00
abstract：:We describe an expression system for high-yield production of recombinant soluble human FasL (rsh- FasL) in CHO cells. After one round of selection for gene amplification, cell lines producing rsh-FasL up to 60 microg/L x 10(6) cells in 24 h were obtained. Cell lines were grown in protein-free medium as suspension cul...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1385/MB:23:3:189
更新日期：2003-03-01 00:00:00
abstract：:NK cells have been seen as potential agents in adoptive immunotherapy for cancer. The main challenge for the success of this approach is to obtain a great quantity of activated NK cells for adoptive transfer. The present study had aimed to evaluate the effect of a feeder layer of irradiated MSCs in the in vitro expans...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-020-00250-2
更新日期：2020-05-01 00:00:00
abstract：:Surface plasmon resonance (SPR) biosensor is a promising technology for its various advantages including the real-time measurement of biomolecular interactions without labeling. A method of hybridizing RNAs on the surface of the streptavidin-coated (SA) sensor chip to study RNA-protein interactions was described in th...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-008-9066-3
更新日期：2008-09-01 00:00:00
abstract：:Molecular biological methods that use antibodies and nucleic acids to detect specific foodborne bacterial pathogens were scarcely known a decade and a half ago. Few scientists could have predicted that these tools of basic research would come to dominate the field of food diagnostics. Today, a large number of cleverly...
journal_title：Molecular biotechnology
pub_type： 杂志文章,评审
doi：10.1007/BF02740817
更新日期：1997-06-01 00:00:00
abstract：:N-Succinyl-amino acid racemase (NSAAR), long referred to as N-acyl- or N-acetyl-amino acid racemase, is an enolase superfamily member whose biotechnological potential was discovered decades ago, due to its use in the industrial dynamic kinetic resolution methodology first known as "Acylase Process". In previous works,...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-015-9839-4
更新日期：2015-05-01 00:00:00
abstract：:Many phytopathogenic micro-organisms such as bacteria and fungi produce pectin methylesterases (PME) during plant invasion. Plants and insects also produce PME to degrade plant cell wall. In the present study, a thermostable pectin methylesterase (CtPME) from Clostridium thermocellum belonging to family 8 carbohydrate...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-017-9997-7
更新日期：2017-05-01 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：: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：:The filamentous fungus Sclerotinia sclerotiorum produces a complete set of cellulolytic enzymes needed for efficient solubilization of native cellulose, the major component of plants. In this work, we reported the molecular characterization of an important glycosyl-hydrolase enzyme classified as endo-β-1,4-glucanase. ...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-013-9714-0
更新日期：2014-04-01 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：: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：: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：:In this paper, we report cloning of a pectate lyase gene from Bacillus amyloliquefaciens S6 (pelS6), and biochemical characterization of the recombinant pectate lyase. PelS6 was found to be identical with B. subtilis 168 pel enzyme with 100% amino acid sequence homology. Although these two are genetically very close, ...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-019-00194-2
更新日期：2019-09-01 00:00:00
abstract：:Megaprimer-based methodology has been widely applied in site-directed mutagenesis, but rarely used in gene splicing. In this article, we describe a modification of the megaprimer PCR method, which can efficiently create and amplify a specific ligated chimeric gene segment in a PCR reaction and under a common PCR progr...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-008-9078-z
更新日期：2008-11-01 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：:A new lipase gene designated as SlLipA was isolated from Serratia liquefaciens S33 DB-1 by the genomic-walking method. The cloned gene contained an open reading frame (ORF) of 1,845 bp encoding 615 amino acids with a conserved GXSXG motif. Genome sequence analysis showed that an aldo/keto reductase gene closed to the ...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-007-9007-6
更新日期：2008-02-01 00:00:00
abstract：:Expression of certain transgenes from an adenovirus vector can be deleterious to its own replication. This can result in the inhibition of virus rescue, reduced viral yields, or, in the worst case, make it impossible to construct a vector expressing the inhibiting transgene product. A gene regulation system based on t...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/BF02686012
更新日期：2007-03-01 00:00:00
abstract：:The REDOX-RESPONSIVE TRANSCRIPTION FACTOR 1 (RRTF1) gene encodes a member of the ERF/AP2 transcription factor family involved in redox homeostasis. The RRTF1 gene shows tissue-specific responsiveness to various abiotic stress treatments including a response to salt stress in roots. An interesting feature of this respo...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-019-00155-9
更新日期：2019-04-01 00:00:00
abstract：:Gene reporter systems play a key role in gene expression and regulation studies. This review describes the ideal reporter systems, including reporter expression vector design. It summarizes the many uses of genetic reporters and outlines the currently available and commonly used reporter systems. Each system is descri...
journal_title：Molecular biotechnology
pub_type： 杂志文章,评审
doi：10.1385/MB:13:1:29
更新日期：1999-11-01 00:00:00
abstract：:Transgenic rice seed can be utilized as a bioreactor to produce high-value recombinant proteins. Mouse interleukin 4 (mIL-4) and mIL-6 were specifically expressed as secretory proteins in rice endosperm by ligating the N-terminal glutelin B-1 (GluB-1) signal peptide and the C-terminal KDEL endoplasmic reticulum retent...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-016-9920-7
更新日期：2016-04-01 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：:Pasteurella multocida is the main cause of haemorrhagic septicaemia (HS) outbreak in livestock, such as cattle and buffaloes. Conventional vaccines such as alum-precipitated or oil-adjuvant broth bacterins were injected subcutaneously to provide protection against HS. However, the immunity developed is only for short ...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-020-00244-0
更新日期：2020-05-01 00:00:00
abstract：:D-Allulose is a rare monosaccharide that exists in extremely small quantities in nature, and it is also hard to prepare at a large scale via chemical or enzyme synthetic route due to low conversion and downstream separation complexity. Using D-psicose epimerase and L-rhamnulose kinase, a method enabling high conversio...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-019-00174-6
更新日期：2019-06-01 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：:Alternaria blight caused by Alternaria brassicicola (Schwein.) Wiltshire and A. brassicae (Berk.) Sacc., is one of the most important disease of rapeseed-mustard, characterized by the formation of spots on leaves, stem, and siliquae with premature defoliation and stunting of growth. These two species are very difficul...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-014-9784-7
更新日期：2014-11-01 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
