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Gene Transfer into Pig and Goat Fetal Fibroblasts by Co-transfection of tPA Transgene and $Neo^r$ Gene
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  • Gene Transfer into Pig and Goat Fetal Fibroblasts by Co-transfection of tPA Transgene and $Neo^r$ Gene
  • Gene Transfer into Pig and Goat Fetal Fibroblasts by Co-transfection of tPA Transgene and $Neo^r$ Gene
저자명
Kim. Bae-Chul,Han. Rong-Xun,Kim. Myung-Yoon,Shin. Young-Min,Park. Chang-Sik,Jin. Dong-Il
간행물명
Reproductive & developmental biology
권/호정보
2009년|33권 2호|pp.107-111 (5 pages)
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한국동물번식학회
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정기간행물|ENG|
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이 논문은 한국과학기술정보연구원과 논문 연계를 통해 무료로 제공되는 원문입니다.
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The transfection efficiency of a transgene into pig and goat fetal fibroblast cells (PFF and GFF, respectively) was tested using co-transfection of a human tissue-type plasminogen activator (tPA) transgene and neomycin-resistant ($Neo^r$) gene, followed by G418 selection. To initially test G418 resistance, GFF and PFF were incubated in culture medium containing different concentration of G418 for 2 weeks, and cell survival was monitored over time. Based on the obtained results, the concentrations chosen for G418 selection were 800 ug/ml and 200 ug/ml for GFF and PFF, respectively. For co-transfection experiments, the pBC1/tPA and $Neo^r$ vectors were co-transfected into GFF and PFF ($1{ imes}10^6$ cells in each case) using the FuGENE6 transfection reagent, and resistant colonies were obtained following 14 days of G418 selection. We obtained 96 and 93 drug-resistant colonies of GFF and PFF, respectively, only 54 and 39 of which, respectively, continued proliferating after drug selection. PCR-based screening revealed that 23 out of 54 analyzed GFF colonies and 5 out of 39 analyzed PFF colonies contained insertion of the tPA gene. Thus, the experimentally determined transfection efficiencies for tPA gene co-transfection with the $Neo^r$ gene were 42.6% for GFF and 12.8% for PFF. These findings suggest that co-transfection of a transgene with the $Neo^r$ gene can aid in the successful integration of the transgene into fetal fibroblast cells.