담배 callus (Nicotiana tabacum L., var Virginia 115)로 부터 guanidium thiocyanate 및 guanidine HCl 방법을 이용 total cellular RNA를 분리 하였으며 oligo (dT)-cellulose 친화성 크로마토그라피 방법에 의해 $poly(A)^+$ RNA를 분리하였다. Protein-synthesizing cell-free system을 이용한 peroxidase mRNA의 translation을 위 해 rabbit reticulocyte lysate의 peroxidase mRNA에 대한 최적 translation 조건을 조사한 결과 2mM $Mg^{2+}$, 80 mM $K^+$, $15;{mu}M$ hemine 및 $1;{mu}g$의 $poly(A)^+$ RNA 일때 최대의 방사능 유입과 peroxidase 활성을 나타냈다. 그러나 어떤 peroxidase population에 있어서는 1 mM $Mg^{2+}$을 최적 조건으로 갖는 경우도 있었다. 6M urea agarose electrophoresis에 의해 total cellular RNA를 분석해 본 결과 16S 및 23S 부근에서 peroxidase mRNA를 확인할 수 있었다. 또한 Anti isoperoxidase $C_3$ 및 $A_4$antiserum을 이용한 immunoprecipitation에 의해 in vitro translation products 내에서 anti $C_3$와 crossreactivity를 갖는 isoperoxidase $C_3$, $C_4$, $C_1/C_2$ 및 anti $A_4$와 crossreactivity를 갖는 isoperoxidase $A_4$, $A_1$을 확인할 수 있었다.
Total celluar RNA was isolated from tobacco callus (Nicotiana tabacum L., var Virginia 115) with guanidium thiocynate and guanidine HCl method. The total RNA was further fractionated by oligo (dT)-cellulose chromatography. For translation of peroxidase mRNAs in cell-free protein-synthesizing system, rabbit reticulocyte lysate was tested and the optimum translational conditions of peroxidase mRNAs were established. The lysate containing 2 mM $Mg^{2+}$, 80 mM $ K^+$ and $15;{mu}M$ hemine was found to be the optimum condition for the translation of one population of peroxidase mRNAs. On the other hand, 1 mM $Mg^{2+}$ was the optimum for the other population of peroxidase mRNAs. The incorporation of [$^{35}S$]-Met into peroxidase was increased up to $1;{mu}g$ of $poly(A)^+$ RNA and then it was decreased. Size fractionation of total cellular RNA was carried out by 6M urea agarose electrophoresis. Peroxidase mRNAs were distributed at slightly above 16S and 23S regions. As a result of immunoprecipitation of in vitro translation products with anti isoperoxidase $C_3$ and $A_4$ antiserum, we found $C_3$, $C_1$ or/and $C_2$, $C_4$ crossreactive to anti $C_3$ antiserum and $A_4$, $A_1$ to anti $A_4$antiserum.
