Abstract:
:The neomycin-resistant gene (neo(r)) is probably the most commonly used selectable marker gene in gene targeting and gene transfection research. In this study, the neo(r) gene construct was introduced into in vitro cultured goat foetal fibroblast cells (IV-5), and the cells were selected with 900 microg/ml G418. The G418-resistant colonies were analysed by neo-specific PCR, karyotyping and anti-intermediate filament proteins antibody (anti-vimentin) staining. Cell cycle analysis of the neo(r) positive foetal fibroblast cell colony (IV-5.1) cultured in a variety of cell cycle-arresting medium indicated that 74.2% of cells cultured in serum-deprived medium for 3 days and 71.7% of cells grown to confluence were at G0/G1 stage of cell cycle, respectively, in comparison to 61.6% of cells in normal culture (cycling) medium. Nocodazole treatment for 17 hr in vitro culture could increase the number of cells at G2/M stage of cell cycle from 20.3% (in cycling medium) to 39.7%. In total, one early pregnancy was observed by B ultra-sound scanning in a surrogate transferred with cloned embryos from IV-5.1 cells at M stage (cells were cultured in nocodazole medium). Seven cloned goats, including two that miscarried at a late stage, were derived from the IV-5.1 cell clone cultured in starved medium (G0). Indeed, one surrogate receiving three blastocysts reconstituted from the starved donor cells, gave birth to three live cloned goats, all of which are healthy and doing well. PCR, Southern blot and G418 resistance in vitro of fibroblast cells from cloned goats confirmed that all cloned goats are positive for neo(r) transgene. This study demonstrates that a foreign gene, such as the neo-resistant gene, can be introduced into goat foetal fibroblast cells, and that the resulting transgenic cells are capable of being cloned to produce 100% transgenic animals.
journal_name
Mol Reprod Devjournal_title
Molecular reproduction and developmentauthors
Zou X,Wang Y,Cheng Y,Yang Y,Ju H,Tang H,Shen Y,Mu Z,Xu S,Du Mdoi
10.1002/mrd.1143keywords:
subject
Has Abstractpub_date
2002-02-01 00:00:00pages
164-72issue
2eissn
1040-452Xissn
1098-2795pii
10.1002/mrd.1143journal_volume
61pub_type
杂志文章abstract::To improve in vitro maturation and to understand the mechanism for meiotic resumption of oocytes, meiotic progression, and its control by hypoxanthine (HX) were studied in goat oocytes. Ovaries were obtained from a local abattoir, and cumulus-oocyte complexes (COCs) and follicular fluid were collected from follicles o...
journal_title:Molecular reproduction and development
pub_type: 杂志文章
doi:10.1002/mrd.10355
更新日期:2003-11-01 00:00:00
abstract::The frameworks (key mechanisms) of the self/non-self-discrimination systems that are found in various organisms have not been actively selected for, but have evolved by genetic drift such that the genetic frequency of random, advantageous mutations has increased within the genomes of these species by natural selection...
journal_title:Molecular reproduction and development
pub_type: 杂志文章
doi:10.1002/mrd.22134
更新日期:2013-01-01 00:00:00
abstract::Dickkopf-like 1 (DkkL1) is related to the Dickkopf gene family, a group of proteins that are characterized as secreted antagonists of Wingless (Wnt) signal transduction proteins. DkkL1 mRNA is found in preimplantation mouse embryos and in developing neural tissue, but in adults it is found primarily in the testes. In ...
journal_title:Molecular reproduction and development
pub_type: 杂志文章
doi:10.1002/mrd.20314
更新日期:2005-08-01 00:00:00
abstract::We aimed in this study to explore if sphingosine-1-phosphate (S1P) reduces apoptosis of primordial follicles during cryopreservation of human ovarian cortical samples. Ovarian cortical tissue fragments obtained from young patients who underwent laparoscopic excision of benign ovarian cysts were used for the experiment...
journal_title:Molecular reproduction and development
pub_type: 杂志文章
doi:10.1002/mrd.23043
更新日期:2018-11-01 00:00:00
abstract::Oocyte maturation is accompanied by differentiation of surrounding cumulus cells. These cells produce hyaluronic acid (HA) and its storage in intercellular spaces results in expansion of the cells. The cumulus cells also accumulate cyclooxygenase-2 (cox-2) during maturation. Both expansion and cox-2 storage are regula...
journal_title:Molecular reproduction and development
pub_type: 杂志文章
doi:10.1002/mrd.10265
更新日期:2003-05-01 00:00:00
abstract::In vitro embryo production is an established method for both humans and animals, but is fraught with inferior development and health issues in offspring born after in vitro fertilization procedures. Analysis of epigenetic changes caused by exposure to in vitro conditions should shed light on potential sources of these...
journal_title:Molecular reproduction and development
pub_type: 杂志文章
doi:10.1002/mrd.22922
更新日期:2017-12-01 00:00:00
abstract::The aim of the present study was to investigate the mechanism of androgen-mediated, prolonged sperm-storage in the female reproductive tract of the bat, Scotophilus heathii. The bat treated in vivo with flutamide, an androgen antagonist, showed loss of spermatozoa at the storage site, utero-tubal junction. Immunocytoc...
journal_title:Molecular reproduction and development
pub_type: 杂志文章
doi:10.1002/mrd.21328
更新日期:2011-07-01 00:00:00
abstract::Basigin, a transmembrane glycoprotein belonging to the immunoglobulin superfamily, has been shown to be essential for fertilization and implantation. The aim of this study was to determine the expression and hormonal regulation of basigin gene in mouse uterus during the peri-implantation period. Basigin immunostaining...
journal_title:Molecular reproduction and development
pub_type: 杂志文章
doi:10.1002/mrd.10128
更新日期:2002-09-01 00:00:00
abstract::PLAC1 is a recently described, trophoblast-specific gene that localizes to a region of the X-chromosome important in placental development. Immunohistochemical analysis demonstrated that PLAC1 polypeptide localizes to the differentiated syncytiotrophoblast throughout gestation (8-41 weeks) as well as a small populatio...
journal_title:Molecular reproduction and development
pub_type: 杂志文章
doi:10.1002/mrd.20272
更新日期:2005-07-01 00:00:00
abstract::Under the influence of gonadotropins or growth factors, a close cooperation develops between cumulus cells and the oocyte that is implicated in transmitting signals involved in maintaining or releasing the meiotic arrest in the oocyte. While cyclic adenosine 5'-monophosphate (cAMP) is a key molecule in maintaining the...
journal_title:Molecular reproduction and development
pub_type: 杂志文章,评审
doi:10.1002/mrd.21332
更新日期:2011-10-01 00:00:00
abstract::Hyperactivated motility was studied in guinea pig spermatozoa. In the presence of the local anesthetic procaine, a high number of sperm cells (64%) showed hyperactivation when incubated in minimal culture medium with pyruvate, lactate, and glucose. Hyperactivated motility was dependent on glucose in the medium. Sperm ...
journal_title:Molecular reproduction and development
pub_type: 杂志文章
doi:10.1002/mrd.1080380309
更新日期:1994-07-01 00:00:00
abstract::MAP kinase (MAPK) and its activator, MAP kinase kinase (MAPKK), are commonly activated by a variety of extracellular stimuli in mammalian cells and in the process of Xenopus oocyte maturation. In order to investigate the function of the MAPK cascade in oocyte maturation, we produced an anti-Xenopus MAPKK which specifi...
journal_title:Molecular reproduction and development
pub_type: 杂志文章,评审
doi:10.1002/mrd.1080420417
更新日期:1995-12-01 00:00:00
abstract::Testis induction is associated with gonadal Sry and Sox9 expression in mammals, and with Sox9 expression in vertebrates where Sry is absent. In mammals, Sry might initiate testis induction by upregulating Sox9 expression; however, direct evidence supporting this hypothesis is lacking. Models of Sry-negative XX sex rev...
journal_title:Molecular reproduction and development
pub_type: 杂志文章
doi:10.1002/mrd.10317
更新日期:2003-08-01 00:00:00
abstract::We determined the incidence of activation, male pronuclear formation, and apposition of pronuclei in porcine oocytes following intracytoplasmic injection of various porcine sperm components and foreign species spermatozoa, such as that of cattle, mouse or human. The porcine oocytes were activated by injection of a spe...
journal_title:Molecular reproduction and development
pub_type: 杂志文章
doi:10.1002/(SICI)1098-2795(199905)53:1<84::AID-MRD10>
更新日期:1999-05-01 00:00:00
abstract::We report use of an in vitro assay (Barbato et al., 1998: Biol Reprod 58:686-699) to assess binding ability of cauda epididymal mouse sperm to a surrogate zona pellucida and effect of a synthetic peptide (Amann et al., 1999: J Androl 20: 42-46) on fertilization ability in in vitro fertilization (IVF) tests. Sperm from...
journal_title:Molecular reproduction and development
pub_type: 杂志文章
doi:10.1002/1098-2795(200012)57:4<406::AID-MRD13>3.0.C
更新日期:2000-12-01 00:00:00
abstract::Transcription factors inhibit or assist RNA polymerases in the initiation and maintenance of transcription; however, the cell specific expression and roles of transcription factors within bovine ovarian follicles during development are unknown. The aim of present study was to determine if the expression of transcripti...
journal_title:Molecular reproduction and development
pub_type: 杂志文章
doi:10.1002/mrd.20819
更新日期:2008-05-01 00:00:00
abstract::The reproductive tract environment at conception programs the developmental trajectory of the embryo, sets the course of pregnancy, and impacts offspring phenotype and health. Despite the fundamental importance of this stage of reproduction, the rate-limiting regulatory mechanisms operating locally to control fertilit...
journal_title:Molecular reproduction and development
pub_type: 杂志文章,评审
doi:10.1002/mrd.22823
更新日期:2017-09-01 00:00:00
abstract::Limited understanding of the importance of known pluripotency factors in pig embryonic stem cells (ESC) impedes the establishment and validation of porcine ESC lines. This study evaluated the expression of known mouse ESC and human ESC (hESC) pluripotency markers in in vivo inner cell mass (ICM) and in vitro-cultured ...
journal_title:Molecular reproduction and development
pub_type: 杂志文章
doi:10.1002/mrd.20780
更新日期:2008-03-01 00:00:00
abstract::Mouse oocyte development in vitro has been studied in the past several years, but no evidence showed that the fertilizable oocytes could be obtained from the fetal mouse germ cells before the formation of the primordial follicles. In this study, an efficient and simple method has been established to obtain the mature ...
journal_title:Molecular reproduction and development
pub_type: 杂志文章
doi:10.1002/mrd.20535
更新日期:2006-10-01 00:00:00
abstract::The mouse metallothionein (MT) gene family consists of four known members (MT-I through IV) clustered on chromosome 8. Studies reported herein examine the expression and regulation of the MT-III and MT-IV genes in specific cell types in the maternal reproductive tract, developing embryo, and fetus known to express the...
journal_title:Molecular reproduction and development
pub_type: 杂志文章
doi:10.1002/(SICI)1098-2795(199601)43:1<25::AID-MRD4>3
更新日期:1996-01-01 00:00:00
abstract::The present study was conducted to reveal effects of in ovo injection of nonsteroidal aromatase inhibitor (Fadrozole) or estradiol at day 3 of incubation on mRNA levels of P45017alphahydroxylase (P450c17), P450 aromatase (P450arom) and anti-Müllerian hormone (AMH) in the chicken gonads. The mRNA levels in the gonads a...
journal_title:Molecular reproduction and development
pub_type: 杂志文章
doi:10.1002/(SICI)1098-2795(200001)55:1<20::AID-MRD4>3
更新日期:2000-01-01 00:00:00
abstract::Altering incubation temperature during embryogenesis has an impact on chicken embryo growth, but the underlying molecular mechanisms are not understood; the present study was performed to address these changes. Broiler eggs were incubated at low (36.8°C), control (37.8°C), and high (38.8°C) temperatures between Embryo...
journal_title:Molecular reproduction and development
pub_type: 杂志文章
doi:10.1002/mrd.22596
更新日期:2016-01-01 00:00:00
abstract::In the equine, the zona pellucida (ZP) is the major barrier to successful in vitro fertilization. Therefore the aim of our studies was to analyze species-specific features of the equine ZP in regard to structure and glycoprotein ZPB and ZPC expression sites during oocyte development and embryogenesis. The equine ZP re...
journal_title:Molecular reproduction and development
pub_type: 杂志文章
doi:10.1002/mrd.20501
更新日期:2007-07-01 00:00:00
abstract::Configuration of germinal vesicle (GV) chromatin has been studied and found correlated with the developmental competence of oocytes in several mammalian species. A common feature in the configuration of GV chromatin in the species studied so far is that the diffuse chromatin (the so called "NSN" pattern) condenses int...
journal_title:Molecular reproduction and development
pub_type: 杂志文章
doi:10.1002/mrd.20251
更新日期:2005-06-01 00:00:00
abstract::Embryonic stem (ES) cell lines are routinely derived from in vivo produced blastocysts. We investigated the efficiency of ES cells derivation from in vitro produced blastocysts either in monoculture or sequential culture. Zygotes from hybrid F1 B6D2 mice were cultured in vitro to the blastocyst stage in Potassium (K(+...
journal_title:Molecular reproduction and development
pub_type: 杂志文章
doi:10.1002/mrd.20512
更新日期:2006-08-01 00:00:00
abstract::These experiments were designed to test the effects of an electrofusion and an electroporation pulse on bovine sperm-hamster egg development. In experiment 1, single motile sperm were injected into the perivitelline space of each egg. A 4,500 V/cm, 30 microseconds fusion pulse (FP) was applied while sperm-egg membrane...
journal_title:Molecular reproduction and development
pub_type: 杂志文章
doi:10.1002/mrd.1080270212
更新日期:1990-10-01 00:00:00
abstract::Insulators are located at the boundaries of differentially regulated genes and delimit their interactions by establishing independent chromatin structures. Recently, an insulator sequence has been found in the 5'-flanking region of arylsulfatase (ARS) gene from sea urchin. To investigate functional conservation of thi...
journal_title:Molecular reproduction and development
pub_type: 杂志文章
doi:10.1002/1098-2795(200011)57:3<232::AID-MRD4>3.0.CO
更新日期:2000-11-01 00:00:00
abstract::The gonadotropin releasing hormone (GnRH) has long been considered as a neuropeptide involved in the control of the reproductive cycle. However, the presence of GnRH and its receptors in various tissues, including ovary and testis, suggests a role as autocrine/paracrine factor. In the present study, we report the expr...
journal_title:Molecular reproduction and development
pub_type: 杂志文章
doi:10.1002/mrd.1000
更新日期:2001-05-01 00:00:00
abstract::In studies of somatic cell nuclear transfer (SCNT), the ability of factors within the oocyte to epigenetically reprogram transferred nuclei is essential for embryonic development of the clone to proceed. However, irregular patterns of X-chromosome inactivation, abnormal expression of imprinted genes, and genomic DNA h...
journal_title:Molecular reproduction and development
pub_type: 杂志文章
doi:10.1002/mrd.21306
更新日期:2011-05-01 00:00:00
abstract::In pigs, the morphology and cell number of in vitro-produced blastocysts are inferior to those of their in vivo counterparts. The objective of this study was to increase developmental competence and to gain an understanding of cell allocation in blastocysts derived from the aggregation of four-cell stage porcine embry...
journal_title:Molecular reproduction and development
pub_type: 杂志文章
doi:10.1002/mrd.20728
更新日期:2007-11-01 00:00:00