Publications by authors named "G Speksnijder"

Article Synopsis
  • Chicken blastodermal cells can be cultured briefly and still contribute to various tissues in gamma-irradiated embryos, but creating transgenic birds has been challenging due to low integration rates of transgenes into their genomes.* -
  • This study developed two methods to successfully isolate and culture blastodermal cells with integrated transgenes from specific chicken embryos, using chick embryo extract to promote cell growth.* -
  • Although injected into irradiated embryos resulted in chicks with donor cell tissues and detectable transgene sequences in sperm DNA, breeding these chimeric birds did not lead to successful germline transmission of the transgene.*
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We have previously described the expression of a bacterial protein in the egg white of transgenic chickens using a replication-deficient retroviral vector. Here we report the expression of a glycosylated human protein, interferon alpha-2b (hIFN), in the egg white of transgenic hens. The hIFN secreted into the egg white was biologically active as determined by a viral inhibition assay.

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Using a replication-deficient retroviral vector based on the avian leukosis virus (ALV), we inserted into the chicken genome a transgene encoding a secreted protein, beta-lactamase, under the control of the ubiquitous cytomegalovirus (CMV) promoter. Biologically active beta-lactamase was secreted into the serum and egg white of four generations of transgenic chickens. The expression levels were similar in successive generations, and expression levels in the magnum of the oviduct were constant over at least 16 months in transgenic hens, indicating that the transgene was stable and not subject to silencing.

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We have developed a novel method of DNA extraction combined with a high-throughput method of gene detection allowing thousands of potentially transgenic chicks to be screened quickly and reliably. By using this method and a replication-deficient retroviral vector based on avian leukosis virus (ALV), we have demonstrated germline transmission of three different transgenes. Several generations of chickens carrying intact transgenes were produced, validating the use of the ALV retroviral vectors for large-scale production of transgenic flocks.

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Three experimental strains of breeder chickens were accidentally exposed to Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS), presumably from a newly introduced group of leghorn-type pullets. The experimental strains subsequently became infected and were diagnosed positive for MG and MS by the serum plate agglutination (SPA) test and confirmed by the hemagglutination inhibition (HI) test and the polymerase chain reaction (PCR) of tracheal swabs. Treatment with 10 mg/kg enrofloxacin via drinking water for 14 days was elected.

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