Studies were carried out on the preparation procedure and characterization of the ribosome of Mycobacterium 607 (M. 607).
The microbe was cultured with the tween 80 synthetic medium at 37°C for four days. Then, the cells were harvested by centrifugation, and washed with and resuspended in the 0, 01 M tris buffer containing 0, 01 M of MgCl
2 (tris-Mg buffer). The cell free extract of the washed cells was prepared by passing through the French pressure cell at 400 Kg/cm
2 followed by centrifugation at 5, 000 g for 20 minutes.
Particulate fractions with various sizes were isolated by successive ultracentrifugation at 20, 000 g for 60 minutes, 60, 000 g for 90 minutes, and 105, 000 g for 120 minutes. These particulate fractions and final supernatant were designated as 20p60, 60p90, 105p120 and 105s120 fractions. Each particulate fraction was suspended in the tris-Mg buffer, and chemical and physical analysis was performed. Results obtained were as follows:
1) It was demonstrated that the 60p90 and 105p120 fractions contain high nucleic acid as measured U-V spectrophotometorically.
2) The 60p90 and 105p120 fractions consisted of 94, 1 S, 74, 4 S, 45, 6 S particles and 74, 4 S, 45, 6 S particles, respectively.
3) The RNA protein ratios of 60p90 and 105p120 fractions were 56, 3: 43, 7, 46, 6: 53, 4, respectively.
4) Furthermore, it was shown that both 60p90 and 105p120 fractions could incorporate
14C-amino acids into their protein fractions in the presence of 105s120 fraction and ATP and its generator.
These results indicate that the 60p90 and 105p120 fractions may be the ribosome fractions of M. 607.
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