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Contribution on the biochemical investigation of external factors required for type transformation in thePneumococcus

Дополнения к виохимическому иэучению впешних ϕакторов, неовходимых для трансϕормации типа у пневмококка

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Summary

  1. (1)

    It was found that different anti-R-Pneumococcus sera display a great variability in DNase activity contained therein. There are sera in which a high DNase activity remains even after inactivation at 65° C for 30 min. At dilutions which are commonly used in the experiments a certain residual activity is always found.

  2. (2)

    The presence of suitable concentrations of salts with univalent cations in the transformation medium inhibits the DNase activity in it, thus regularizing the results obtained during type transformation in the Pneumococcus. This explains the results of our earlier work concerning the significance of these salts in the transformation reaction even if their role is manifold.

  3. (3)

    By preliminary estimation of DNase activity in sera or serous fluids, globulin and albumin preparations, it can readily be shown whether these sera are or are not suitable for supporting the transformation reaction. Antisera with a high residual activity of DNase do not support the transformation reaction unless DNase had been suitably inhibited.

  4. (4)

    High DNase content in serum may by inhibited by higher temperature or occasionally by increased salt concentration. By using a mixture of citrate, magnesium ions and a small amount of pancreatic DNase activated for 15 min. at 30° C and then inactivated for 15 min. at 60° C, together with DNA, a positive result may be achieved during transformation using some antisera with a high DNAse activity which will not allow such a reaction to proceed without the agents added.

  5. (5)

    The transformation reaction always takes place in the presence of a small amount of DNase which is blocked by different factors in the system. The possible active role of DNase in the transformation reaction is discussed. The author is indebted to Academician I. Málek for the interest with which he followed the entire work described. Technical assistance of PhMr. P. Křečková and Mr. J. Kubíček is acknowledged. Our thanks are due to Dr. Hilgert of the Institute of Experimental Biology and Genetics for a sample of DNA from chick erythrocytes.

Abstract

  1. (1)

    Мв установили, что раэличные анти-R пневмококковые сыворотки отличаются эначителыюй иэменчивостъю активности содержащейся в них деэоксирибонуклеаэ ы(ДН аэа). Это сывороеки, в которых высокая степенъ активности ДНϕэы сохраняется и после инактивирования при 65° C в течение 30 мин. В раэведениях, применяемых обычно для опытов, всегда находится иэвестная остаточная активностъ ДНаэы.

  2. (2)

    Присутствие в трансϕормационной среде соответствующих концентраций солей с одновалентными катионами подавляет активностъ ДНаэы, чем определяются реэулътаты трансϕормации типа пневмококка. Этим объясняются реэулътаты наших предшествовавших работ по иэучению эначения этих солей для реакдии трансϕормадии, хотя ролъ этих солеи, по-видимому, является более многосторонней.

  3. (3)

    Путем предварителъного определения активности ДНаэы в сыворотках или в сероэных жидкостях, глобулино вых и алъбуминовых препаратах можно быстро установитъ, пригодны ли они для поддержания реакции трансϕорма ции - или нет. Антисыворотки с высокой остаточной активностъю ДНϕэы не способствуют реакции трансϕормации, если не имело места предварителъное соответственное угнетение ДНаэы.

  4. (4)

    Угнетение ДНаэы в сыворотках воэможно с помощъю повышения температуры или иногда с помощъю более высоких концентрации солеи. лсполъэуя смесъ цитратов, ионы Mg++и неболъшое количество панкреатическои ДНϕэы, активированной 15 мин. при 30° C и после этого инактивированной 15 мин. при 60° С вместе с ДНК, можно добитъся положителъных реэулътатов трансϕормации с применением некоторых антисывороток с высокой активностъю ДНаэы там, шже беэ этои смеси это невоэможно.

  5. (5)

    Реакция трансϕормации протекает всегда в присутствии неболъшого количества ДНаэы, которая в системе блокируется раэличными ϕакторами. — Обсуждается воэможностъ активной роли ДНаэы в реакции трансϕормации.

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  1. Department of Microbial Genetics and Variability, Institute of Microbiology, Czechoslovak Academy of Sciences, Prague 6

    Margita Kohoutová

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  1. Margita Kohoutová
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Kohoutová, M. Contribution on the biochemical investigation of external factors required for type transformation in thePneumococcus . Folia Microbiol 7, 33–46 (1962). https://doi.org/10.1007/BF02926329

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