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KMID : 0900920030270030197
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Korean journal of Animal Reproduction
2003 Volume.27 No. 3 p.197 ~ p.206
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Transfer and Expression of the Recombinant hFSH Gene in CHO Cells and Transgenic Chickens using Retrovirus Vector System
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Kwon Mo-Seon
Koo Bon-Chul
Shim Ho-Sup
Park Chang-Sik
Lee Seung-Ho
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Abstract
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hFSH (human follicle stimulating hormone) is heterodimer consisting of alpha and beta subunits. Since assembly of the both subunits in the cell is often the rate-limiting step in production of functional hormone, single-chain hormones have been engineered by genetically linking two different cDNA fragments with a linker sequence. Using retrovirus vector system, the resulting recombinant hFSH gene was transferred in CHO cells and chicken embryos, and the expression of the gene was investigated. In CHO cells, protein synthesis from the single-chain FSH gene was 17 fold higher than that from the heterodimeric counterpart. In the study of transgenic chickens, ten of the eleven chicks hatched from 62 embryos manupulated with recombinant retrovirus stock was determined to carry transgenic genes. RT-PCR analyses confirmed transcription of the single-chain FSH gene, however, no recombinant FSH was detected from the blood samples.
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KEYWORD
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hFSH gene, CTP, CHO cell, Transgenic chicken, RT-PCR
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