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KMID : 0900920020260020095
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Korean journal of Animal Reproduction
2002 Volume.26 No. 2 p.95 ~ p.104
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Production of Transgenic Porcine haboring the Human Erythropoietin(EPO) Gene
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Park Jin-Ki
Min Kwan-Sik
Seong Hwan-Hoo
Jeon Ik-Soo
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Abstract
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This study was performed during the four seasons for the production of transgenic pigs containing the human erythropoietin(hEPO) transgene. Purebred Landrace gilts and sows approximately 8∼15 months of age (n=42) were used fur the collection of 1-cell zygotes for DNA microinjection and transfer. Retrospectively, estrus synchronization and superovulation schemes were evaluated to assess practicality for zygote collection. Synchronization and superovulation procedures were used that cyclic gilts were synchronized with 20mg altrenogest (ALT) per day for 9days after PG600 administration followed by superovulation with 1500IU pregnant mares serum gonadotropin (PMSG) and 500IU human chorionic gonadotrophin (hCG). Preparation of recombinant gene for microinjection is mice whey acidic protein promoter (mWAP) linked to human erythropoietin (hEPO) gene. After hormone treatment, 650 embryos were collected from 23 donors and 83.1% (540/650) embryos were in 1-cell stage which can be visualized the pronuclei for DNA microinjection. A total of 543 DNA microinjected embryos fiom donors were transferred to 19 synchronized recipients, seven of them maintained pregnancy and delivered 47 piglets. One of the 47 offsprings were determined to have transgene by PCR analysis. The overall rate of transgenic production was 2.13% (tansgenic/offspring). This study provides the success and useful information regarding production of transgenic pig for bioreactor research.
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KEYWORD
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Transgenic pig, mWAP, Human EPO gene, Microinjection, Superovulation
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