Novel gene transfer into the fertilized eggs of gold fish (Carassius auratus L. 1758)

Abstract

Novelgene transfer into the fertilized eggs of gold fish 31 KREYSZIG, E., 1970: Statistische Methoden und ihre Anwendungen. 422 pp. Vandenhoek und Rupprecht, Gottingen. MCCAULEY, R.; CASSELMAN, J., 1982: Final preferendum of fish -an overestimation of the optimum temperature for growth. Abstract No. 193, 4th Congress of European Ichthyologists, Hamburg TOM, E. 0.; GULYAS, P.; OLAH, J., 1982: Effect of temperature on growth, food conversion and survival of sheatfish (Silurus ghnis L.) and common carp (Cyprinus carpi0 L.) at sublethal ammonia concentrations. Aquacultura Hungarica, Vol. III,51-56. 20.-24.9.1982. SummaryNovel gene which was microinjected into fertilized eggs of the goldfish replicated during the embryogenesis. A proportion of the novel gene has been integrated into the host DNS of the 50-day-old transgenic goldfish. Zusarnmenfassung Ubertragung von Fremdgenen in befruchtete Eier des Goldfsches (Carassius auratus L. 1758)Durch Mikroinjektion in befruchtete Eier des Goldfisches eingebrachte Fremdgene wurden w a r e n d der Embryogenese repliziert. Ein Teil der Fremd-DNS ist in die DNS der 50 Tage alten Goldfische integriert worden. RisumiTransmission des gPnes aliens dans des mufs fertilisis du Cyprin dori (Carassius auratus L. 1758) De genes nouveaux sont microinjectis dans des ceufs fenilisis des embryons de Carassius aurutus. Ces genes sont ripliquis. Un part de la DNS nouveau itait integri dans la DNS des Carassius auratus originaux i ce moment ages 50 journies.In recent years, purified genes have been microinjected into fertilized eggs or embryos of mammals, amphibians, and insects species in order to assess the fate and expression of the novel gene during the development in whole animal system [for review, see (3)]. Results of experiments performed on Xenopus Levis demonstrated that the microinjected, cloned DNA sequences could be replicated and faithfully transcribed during embryogenesis (2,7). PALMITER et al. (1982) (5) introduced into the pronuclei of fertilized eggs of mice a hybrid gene containing the promoter of the mouse metallothionein-1 gene fused to the structural gene for rat growth hormone. Some transgenic mice grew significantly larger than their littermates. Thus it is hoped to use the gene transfer method to develop rapidly growing U.S. A B E D 8 m BPJ -P R 322 -m flanking sequencea and introns m O X O M -hCH flanking sequences and introns 0 hGH cxons Fig. 1.The structure of the novel gene: (A) 9.4 kilobase pair (kb) BPVMG linear sequence as a novel gene to be microinjected into gold fish eggs consists of 69% BPV (bovine papilloma virus) transforming sequence followed by an MG hybrid gene. The MG contains an EcoRl/BglZ fragment of the promoter of the mouse MT-1 (metallothionein-1) gene fused to the BamHl site of a hGH (human growth hormone) mini-gene that includes all the coding sequence and the first of its four intervening sequences. (B) Recombinate plasmid pBPVMG-6 is an MG hybrid gene inserted into a pBR-BPV vector (modified and reconstructed according to PAVLAKIS and HAMER 198...

Retention of the Developmental Pluripotency in Medaka Embryonic Stem Cells after Gene Transfer and Long-term Drug Selection for Gene Targeting in Fish

“…In fish, a transgene is usually delivered directly into * Author for correspondence E-mail: dbshyh@nus.edu.sg eggs by microinjection (Zhu et al, 1985). In most cases the founder animals from injected embryos are highly mosaic for the introduced gene, and desired integration of a transgene and its proper expression has been rarely achieved (Schartl et al, 1998).…”

Enhanced grass carp reovirus resistance of Mx-transgenic rare minnow (Gobiocypris rarus)

“…They have Mx genes of their own but introducing more potent Mx genes might improve their antiviral resistance. Transgenic technology also has the potential to generate fast-growing [30], freeze-resistant [40], and disease-resistant fish [41] for aquaculture. To check the antiviral activity of the GrMx in vivo, we constructed the expression vector for transgenic fish and examined the survival rate after artificial infection.…”

“…The linearized DNA fragment was purified using gel extraction kit (Axygen). After artificial insemination, about 1 nl of linearized recombinant plasmid (100 pg nl À1 ) was microinjected into rare minnow embryo at the 1-2 cell stage under a dissecting microscope with a micromanipulator [28][29][30]. The control group was injected with linearized pCMV-EGFP plasmid.…”

Gene transfer and mutagenesis mediated by Sleeping Beauty transposon in Nile tilapia (Oreochromis niloticus)

“…Since the generation of transgenic goldfish through growth hormone gene transfer has been succeeded (Zhu et al 1985), many of other transgenic fish are successfully constructed, including rainbow trout (Chourrout et al 1986), medaka (Ozato et al 1986), tilapia (Martinez et al 1996), salmon (Fletcher et al 1988), zebrafish (Stuart et al 1988), channel catfish (Dunham et al 1987), and common carp (Zhang et al 1990). Meanwhile, a number of gene transfer approaches including direct microinjection (Zhong et al 2002), retrovirus infection (Linney et al 1999), electroporation (Xie et al 1993), sperm-mediation (Khoo et al 1992) and particle-gun bombardment (Yamauchi et al 2000) were successfully used to deliver foreign DNA into different fish species, but direct microinjection of DNA into the fertilized eggs at one-cell stage has been the prevalent method.…”