TNF-α 유전자로 형질전환된 효모의 성장양태 및 단백질 생산 = Growth and production of protein in transformed yeast with human TNF-α gene

국문 초록 (Abstract)

사람 TNF-a를 생산하도록 GPD promoter와 AG promoter로 형질전환된 S. cerevisiae가 보이는 성장양상, 총분비단백질, pretease 활성을 측정하였다. GPD promoter로 형질전환 된yeast는 63.6 g/L의 성장이 일어났고 총분비단백질은 9 mg/L이며, pretease 활성은 2.55 X10^(4) unit/L 였다. AG promoter로 형질전환된 yeast의 경우엔 68 g/L의 성장이 측정되고 총분비단백질은 27 mg/L, pretease 활성1.55 X10^(4) unit/L였다. 단백질과 protease 분비량은 세포의 성장과 비례하는 것을 알 수 있었고, GPD promoter로 발현되는 TNF-a의 양은 배양초기에는 검출 한계 이하였고 15시간 이후에 증가하는 양상을 보여 배양 후 35시간째에 가장 많은 양 약 2535 ng/ml 정도가 분비되는 것으로 나타났다.

다국어 초록 (Multilingual Abstract)

We measured growth, total secreted protein and total secreted protease of transformed Saccharomyces cerevisiae with human TNF-α gene. Two different promoters, GPD promoter and AG promoter, were used for heterologous expression of human TNF-α gene. The growth which transformed TNF-α:GPD promoter was 63.9 g/L and total secreted protein was 9mg/L, protease activity was 2.55 x 10^(4) unit/L. In another case at TNF-α:AG promoter the growth was 68 g/L, total secreted protein was 27 mg/L and protease activity was 1.55 X 10^(4) unit/L. We could know total secreted protein and protease were directly proportional increase with growth. In initial time of culture, we could not see the TNF-α content in GPD promoter but after 15hours the yeast produced TNF-α. The highest level of product of TNF-a was 2535 ng/ml at 35hours after culture.