Abstract：
:Gene amplification is widely used for the production of pharmaceuticals and therapeutics in situations where a mammalian system is essential to synthesise a fully active product. Current gene amplification systems require multiple rounds of selection, often with high concentrations of toxic chemicals, to achieve the highest levels of gene amplification. The use of these systems has not been demonstrated in specialised mammalian cells, such as embryonic-stem cells, which can be used to generate transgenic animals. Thus, it has not yet proved possible to produce transgenic animals containing amplified copies of a gene of interest, with the potential to synthesise large amounts of a valuable gene product. We have developed a new amplification system, based around vectors encoding a partially disabled hypoxanthine phosphoribosyltransferase (HPRT) minigene, which can achieve greater than 1000-fold amplification of HPRT and the human growth hormone gene in a single step in Chinese hamster-lung cells. The amplification system also works in mouse embryonic-stem cells and we have used it to produce mice which express 30-fold higher levels of human protein C in milk than obtained with conventional transgenesis using the same protein C construct. This system should also be applicable to large animal transgenics produced by nuclear transfer from cultured cell lines.
journal_name
Transgenic Resjournal_title
Transgenic researchauthors
Melton DW,Ketchen AM,Kind AJ,McEwan C,Paisley D,Selfridge Jdoi
10.1023/a:1008951732020subject
Has Abstractpub_date
2001-04-01 00:00:00pages
133-42issue
2eissn
0962-8819issn
1573-9368journal_volume
10pub_type
杂志文章abstract：:This study generated a transgenic zebrafish line Tg(k18:Ptmaa-RFP) with overexpression of Prothymosin alpha type a (Ptmaa) in the skin epidermis. Red fluorescence first appears very weakly in the early stage, become stronger and mainly restricted in the nuclei of the epithelial cells from 3 dpf-larvae to adult fish. H...
journal_title：Transgenic research
pub_type： 杂志文章
doi：10.1007/s11248-009-9350-4
更新日期：2010-08-01 00:00:00
abstract：:Embryonic stem (ES) cells provide a unique tool for cell-mediated gene transfer and targeted gene mutations due to the possibility of in vitro selection of desired genotypes. When selected cells contribute to the germ line in chimaeric embryos, transgenic animals may be generated with modified genetic traits. Though t...
journal_title：Transgenic research
pub_type： 杂志文章
doi：10.1023/a:1015678416921
更新日期：2002-06-01 00:00:00
abstract：:Agriculture productivity is severely affected by soil salinity. One possible mechanism by which plants could survive salt stress is to compartmentalize sodium ions away from the cytosol. In the present work, transgenic buckwheat plants overexpressing AtNHX1, a vacuolar Na(+)/H(+) antiporter gene from Arabidopsis thali...
journal_title：Transgenic research
pub_type： 杂志文章
doi：10.1007/s11248-007-9085-z
更新日期：2008-02-01 00:00:00
abstract：:Use of huge amounts of antibiotics in farm animal production has promoted the prevalence of antibiotic-resistant bacteria, which poses a serious threat to public health. Therefore, alternative approaches are needed to reduce or replace antibiotic usage in the food animal industry. PR-39 is a pig-derived proline-rich a...
journal_title：Transgenic research
pub_type： 杂志文章
doi：10.1007/s11248-018-0084-z
更新日期：2018-10-01 00:00:00
abstract：:This study was performed to qualify goat fetal fibroblast (GFF) cell lines for genetic modification and somatic cell nuclear transfer (SCNT) to produce human lysozyme (hLYZ) transgenic goats. Nine GFF cell lines were established from different fetuses, and the proliferative lifespan and chromosomal stability were anal...
journal_title：Transgenic research
pub_type： 杂志文章
doi：10.1007/s11248-013-9688-5
更新日期：2013-10-01 00:00:00
abstract：:During development in the murine ventricle, there is a switch in myosin heavy chain gene (MyHC) transcription. The beta-MyHC is expressed in the ventricles during foetal development, but is shut down at or around birth, at which time alpha-MyHC transcription is activated. This antithetical switch is thought to be medi...
journal_title：Transgenic research
pub_type： 杂志文章
doi：10.1007/BF01973758
更新日期：1995-11-01 00:00:00
abstract：:Cre/LoxP mediated chromosomal engineering in embryonic stem (ES) cells has a variety of applications, including the creation of model systems for studying aneuploidy. Targeted meiotic recombination (TAMERE) was proposed as a high efficiency in vivo alternative to effect Cre-mediated recombination, in which Cre recombi...
journal_title：Transgenic research
pub_type： 杂志文章
doi：10.1007/s11248-005-0389-6
更新日期：2005-06-01 00:00:00
abstract：:Childhood-onset dermatitis is one of the most common skin disorders in children. Although various mouse models that mirror aspects of dermatitis have become available, there is still a need for an animal model that develops dermatitis in childhood and is more suitable for performing tissue transplantation experiments....
journal_title：Transgenic research
pub_type： 杂志文章
doi：10.1007/s11248-016-9939-3
更新日期：2016-08-01 00:00:00
abstract：:The silk gland of silkworm Bombyx mori, is one of the most important organs that has been fully studied and utilized so far. It contributes finest silk fibers to humankind. The silk gland has excellent ability of synthesizing silk proteins and is a kind tool to produce some useful recombinant proteins, which can be wi...
journal_title：Transgenic research
pub_type： 杂志文章,评审
doi：10.1007/s11248-014-9826-8
更新日期：2014-10-01 00:00:00
abstract：:Japanese morning glory, Ipomoea nil, exhibits a variety of flower colours, except yellow, reflecting the accumulation of only trace amounts of carotenoids in the petals. In a previous study, we attributed this effect to the low expression levels of carotenogenic genes in the petals, but there may be other contributing...
journal_title：Transgenic research
pub_type： 杂志文章
doi：10.1007/s11248-017-0051-0
更新日期：2018-02-01 00:00:00
abstract：:A green fluorescent protein (GFP) cDNA flanked by inverted terminal repeats (ITR) of adeno-associated virus was constructed. The construct sharply improved the efficiency and specificity of the transient expression of genes driven by two general promoters (cytomegalovirus and medaka beta-actin) and one muscle-specific...
journal_title：Transgenic research
pub_type： 杂志文章
doi：10.1023/a:1016671513425
更新日期：2001-08-01 00:00:00
abstract：:Human endogenous retrovirus-R (HERV-R) is one of a full length HERV with a long open reading frame in the env region. The env transcripts are expressed in various human tissues. To investigate the biological role of HERV-R in vivo, we established two lines of transgenic rats carrying a full sequence of HERV-R under co...
journal_title：Transgenic research
pub_type： 杂志文章
doi：10.1023/a:1023381819572
更新日期：2003-06-01 00:00:00
abstract：:Overexpression of a native gene can cause expression of both introduced and native genes to be silenced by posttranscriptional gene silencing (PTGS) mechanisms. PTGS mechanisms rely on sequence identity between the transgene and native genes; therefore, designing genes with mutations that do not cause amino acid chang...
journal_title：Transgenic research
pub_type： 杂志文章
doi：10.1007/s11248-016-9942-8
更新日期：2016-08-01 00:00:00
abstract：:Baculovirus p 35 protein protects cells from apoptotic cell death by inhibiting caspase activation. We have established transgenic mouse lines specifically expressing p 35 in cardiomyocytes, and primary cardiomyocytes isolated from these mice exhibit resistance to staurosporine-induced apoptosis. In a previous study, ...
journal_title：Transgenic research
pub_type： 杂志文章
doi：10.1007/s11248-005-4350-5
更新日期：2005-10-01 00:00:00
abstract：:It is generally believed that too high or low levels of endothelin-1 (ET-1), a strong vasoconstrictor, may be detrimental to animals. Therefore, in order to understand the in vivo function of ET-1, we used a conditional transgenic approach, Cre/loxP recombination system, to generate transgenic mice that over-express E...
journal_title：Transgenic research
pub_type： 杂志文章
doi：10.1023/b:trag.0000026072.01351.10
更新日期：2004-04-01 00:00:00
abstract：:Using the Sleeping Beauty (SB) transposon system, we have developed a simple method for the generation of Xenopus laevis transgenic lines. The transgenesis protocol is based on the co-injection of the SB transposase mRNA and a GFP-reporter transposon into one-cell stage embryos. Transposase-dependent reporter gene exp...
journal_title：Transgenic research
pub_type： 杂志文章
doi：10.1007/s11248-006-9014-6
更新日期：2006-12-01 00:00:00
abstract：:Milk and meat from cattle and buffaloes contribute 45% of the global animal protein supply, followed by chickens (31%), and pigs (20%). In 2016, the global cattle population of 1.0 billion head produced 6.5 billion tons of cows' milk, and 66 million tons of beef. In the past century, cattle breeding programs have grea...
journal_title：Transgenic research
pub_type： 杂志文章
doi：10.1007/s11248-019-00141-6
更新日期：2019-08-01 00:00:00
abstract：:In this study the effects of gene targeting procedures on the early behaviour and morphological development of the resulting offspring have been investigated. Six groups of mice, each having undergone a specific aspect of the biotechnological procedure, (including electroporation, microinjection and/or embryo culture)...
journal_title：Transgenic research
pub_type： 杂志文章
doi：10.1023/a:1012244404020
更新日期：2001-10-01 00:00:00
abstract：:Myostatin is a well-known negative regulator of skeletal muscle growth. Inhibition of myostatin activity results in increased muscle mass. Myostatin propeptide, as a myostatin antagonist, could be applied to promote meat production in livestock such as pigs. In this study, we generated a transgenic mouse model express...
journal_title：Transgenic research
pub_type： 杂志文章
doi：10.1007/s11248-013-9709-4
更新日期：2013-10-01 00:00:00
abstract：:T-DNA activation tagging was used to generate tobacco mutants with increased tolerance to antimicrotubular herbicides and chilling stress. After transformation, protoplast-derived calli were screened for tolerance to treatments that affect microtubule assembly. In one screen mutants with tolerance to aryl carbamates (...
journal_title：Transgenic research
pub_type： 杂志文章
doi：10.1023/a:1025814814823
更新日期：2003-10-01 00:00:00
abstract：:Genomic DNA blot hybridization is traditionally used to demonstrate that, via genetic transformation, foreign genes are integrated into host genomes. However, in large genome species, such as Allium cepa L., the use of genomic DNA blot hybridization is pushed towards its limits, because a considerable quantity of DNA ...
journal_title：Transgenic research
pub_type： 杂志文章
doi：10.1023/a:1016633410041
更新日期：2001-06-01 00:00:00
abstract：:Agrobacterium rhizogenes-transformed sugar beet hairy roots, expressing dsRNA from the Beet necrotic yellow vein virus replicase gene, were used as a novel approach to assess the efficacy of three intron-hairpin constructs at conferring resistance to rhizomania disease. Genetically engineered roots were similar in mor...
journal_title：Transgenic research
pub_type： 杂志文章
doi：10.1007/s11248-010-9364-y
更新日期：2010-10-01 00:00:00
abstract：:Regulatory approvals for deliberate release of GM maize events into the environment have lead to real situations of coexistence between GM and non-GM, with some fields being cultivated with GM and conventional varieties in successive seasons. Given the common presence of volunteer plants in maize fields in temperate a...
journal_title：Transgenic research
pub_type： 杂志文章
doi：10.1007/s11248-009-9250-7
更新日期：2009-08-01 00:00:00
abstract：:A plastid transformation protocol was developed for Lesquerella fendleri, a species with a high capacity for plant regeneration in tissue culture. Transformation vector pZS391B carried an aadA16gfp marker gene conferring streptomycin-spectinomycin resistance and green fluorescence under UV light. Biolistic transformat...
journal_title：Transgenic research
pub_type： 杂志文章
doi：10.1023/a:1022110402302
更新日期：2003-02-01 00:00:00
abstract：:To characterize the activity of the polyomavirus regulatory region, two hybrid marker genes were constructed. In the first construct, the early promoter regulates expression of the CAT gene and the late promoter regulates expression of the lacZ gene. In the second construct, the lacZ gene was placed under the control ...
journal_title：Transgenic research
pub_type： 杂志文章
doi：10.1007/BF01980202
更新日期：1996-11-01 00:00:00
abstract：:We describe a transgenic microalgal Δ9-elongase pathway transformed in both Brassica napus and Arabidopsis thaliana seed resulting in the production of arachidonic acid (ARA). This pathway is noteworthy for both the production of ARA in seed tissue and the low levels of intermediate C20 fatty acids that accumulate. We...
journal_title：Transgenic research
pub_type： 杂志文章
doi：10.1007/s11248-011-9517-7
更新日期：2012-02-01 00:00:00
abstract：:Seed dormancy is an important agronomic trait: low levels can cause premature germination, while too much can inhibit uniform germination. As an approach to controlling the seed dormancy level in crops, we used Triticeae DOG1-like genes as transgenes. DOG1 is an Arabidopsis gene that underlies natural variation in see...
journal_title：Transgenic research
pub_type： 杂志文章
doi：10.1007/s11248-014-9800-5
更新日期：2014-08-01 00:00:00
abstract：:Phytocystatins are proteinaceous inhibitors of cysteine proteases. They have been implicated in the regulation of plant protein turnover and in defense against pathogens and insects. Here, we have characterized an Arabidopsis phytocystatin family gene, Arabidopsis thaliana phytocystatin 4 (AtCYS4). AtCYS4 was induced ...
journal_title：Transgenic research
pub_type： 杂志文章
doi：10.1007/s11248-013-9735-2
更新日期：2014-02-01 00:00:00
abstract：:Matrix attachment regions (MARs) are DNA elements that can increase and stabilize transgene expression. We investigated the effect of the RB7 MAR on transgenic virus resistance. Constructs for resistance to tomato spotted wilt virus (TSWV) with and without flanking RB7 MARs were used to transform tobacco and produce h...
journal_title：Transgenic research
pub_type： 杂志文章
doi：10.1007/s11248-004-5413-8
更新日期：2005-04-01 00:00:00
abstract：:Several reports of promoters from plants, viral and artificial origin that confer high constitutive expression are known. Among these the CaMV 35S promoter is used extensively for transgene expression in plants. We identified candidate promoters from Arabidopsis based on their transcript levels (meta-analysis of avail...
journal_title：Transgenic research
pub_type： 杂志文章,meta分析
doi：10.1007/s11248-013-9757-9
更新日期：2014-04-01 00:00:00
