Fries minimal 배지와 zinc-nitrogen 결핍 배지에서 각각 배양한 Neurospora crassa로부터 messenger RNA을 얻었다. cell-free reticulocyte system을 이용하여 Neurospora mRNA로부터 in vitro translation 산물을 얻고 면역침전법을 이용하여 이 단백합성 산물내에서 NAD glycohydrolase (EC 3.2.2.5)를 확인하였다. zinc-nitrogen 결핍배지에서의 Neurospora RNA와 mRNA양은 정상배지에서의 양에 비하여 감소하였으나 mRNA의 단백합성능력에는 이상을 보이지 않았다. zinc-nitrogen 결핍배지에서 자란 Neurospora에서 NAD glycohydrolase 특이적 mRNA의 합성을 효소의 비활성도의 증가에 비례하여 증가되지 않은 것으로 보아 NAD glycohydrolase의 증가는 transcription level에서의 조절이 아닌 것으로 사료된다. SDS-polyacrylamide gel 전기영동과 isoelectrofocusing에 의한 단백 합성 산물내의 NAD glycohydrolase 분자량과 pI는 각각 6,000 dalton과 9.0으로서 cell-free reticulocyte system에서의 in vitro translation에서는 post-translational modification이 일어나지 않았음을 시사해 주었다.
The in vitro translation products were obtained by translating the isolated Neurospora messenger RNA from Neurospora crassa cultured in Fries minimal medium and zinc and nitrogen deficient medium and the NAD glycohydrolase (NADase) (EC3.2.2.5) was identified in the translation products by the immunoprecipitation methods. The yields of total RNA and messenger RNA from Neurospora crassa grown in zinc and nitrogen deficient media were lower than those from control media but the function of the synthesis of proteins was not affected. The synthesis of NADase-specific mRNA seemed not to be directly proportional to the increase of the specific activity of the enzyme. The increase of NADase in Neurospora crassa grown in zinc and nitrogen deficient seemed not to be ascribed to the regulation in the level of transcription. The molecular weight and the isoelectric point of NADase in the translational products was around 6,000 dalton and pI 9.0, respectively which were different from the other data obtained from purified Neurospora NADase, suggesting that this translation product was not processed post-translational modification.
