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유전자 도입을 이용한 사람 TGF-β1 발현 세포주 제조
이종원,남상욱,김평현,이주범,전계택 한국유전학회 1999 Genes & Genomics Vol.21 No.3
Transforming Growth Factor-β1 (TGF-β1) is a pleiotrophic polypeptide involved in various biological activities including cell growth, differentiation and extracellular matrix deposition. Since TGF-β1 plays important roles in wound healing processes as well as in immune regulation, TGF-β1 is considered as a potential therapeutic agent for wound healing or organ transplantation. To obtain large quantity of human recombinant TGF-β1, an expression plasmid was constructed and transfected into Chinese hamster ovary (CHO) cells. A TGF-β1 producing clone was selected by measuring TGF-β1 levels in the medium and incubated in increasing concentrations of methotrexate up to 60 μM for the amplification of TGF-β1 cDNA in conjunction with a dihydrofolate reductase (dhfr) gene. The cell line selected in the medium containing 60 μM methotrexate produced TGF-β1 at the concentration of 12 ng/㎖ per 10^5 cells/day, which is approximately 9 fold over that of the transfected cells incubated without methotrexate. The selected cell line showed a slow growth rate and changed morphology. Overall, the established CHO cell line in this study would be useful to produce recombinant human TGF-β1.
멕시코산 Axolotl , Ambystoma mexicanum 의 FGF - 8 cDNA 절편 클로닝
정해문,남상욱,김원선,한만종 한국유전학회 1997 Genes & Genomics Vol.19 No.2
FGF-8 (fibroblast growth factor-8) has been known to play crucial roles in various developmental processes such as initiation, outgrowth and patterning of vertebrate limbs, especially in amniotes. However, it is not known whether FGF-8 plays similar roles in the developmental processes of amphibians. Here, we report a partial fragment cloning of an FGF-8 cDNA from embryo and limb blastema cDNA libraries of a salamander, Mexican axolotl (Ambystoma mexicanum). The 1.27 kb cDNA fragment cloned in the present study contains a coding region covering 192 amino acid residues with conserved consensus amino acid residues of FGF family and shows 85% amino acid identity with that of mouse, and 91% amino acid identity with those of chick and Xenopus. Therefore, the cDNA clone obtained in the present study is believed to be a partial fragment of a putative FGF-8 cDNA of Mexican axolotl.
Partial Cloning of Xhox3.1 Gene in Xenopus laevis
정해문,김태균,남상욱,고제명 한국유전학회 1996 Genes & Genomics Vol.18 No.2
VNTR(variable number tandem repeat) loci in the genome are characterized-as their copy number of the same repeat unit and are extremely polymorphic having potentially hundreds of alleles at a single locus. In this study, allele distribution on the hTPO VNTR marker was analyzed by PCR(polymerase chain reaction) in Korean population(100 individuals). Twenty two alleles were identified in the hTPO VNTR locus and alleles 7(8.5%), 10(11.0%), and 15(15.0%) showed three highest peaks. The allele frequency of hTPO VNTR locus is in the 95% confidence interval. PIC(polymorphism information content) value calculated on hTPO VNTR loci was 0.92. POE(power of exclusion) value used in paternity cases was 0.80. MP(matching probability) used in forensic identification was 0.018. In conclusion, statistical data from hTPO VNTR locus showed this locus is a very useful genetic marker for the linkage study, evolutionary genetics and forensic application.
Xenopus Oocyte 에 존재하는 Fibroblast Growth Factor - Like cDNA 의 부분적 클로닝과 발현 특성 분석
김현희,정해문,남상욱,김원선,고제명 한국유전학회 1997 Genes & Genomics Vol.19 No.3
We cloned a partial sequence of novel Xenopus cDNA which showed a significant homology to fibroblast growth factor cDNA. This gene was expressed maternally and presented throughout embryonic stages. The level of this gene expression was gradually decreased during the developmental stages from fertilized egg to early gastrula and began to increase after mid gastrula stage, indicating zygotical expression of the gene after mid gastrula. The wholemount in situ hybridization analysis revealed that this novel gene was expressed in somites, notochord, central nervous system, eye, gill slit and cement gland. These expression analyses indicated that this novel gene might be involved in neuronal development or formation of chordamesoderm.