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Regulation of the lactose operon in vitro

I. Transcription of the lactose operon

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Summary

Using DNA of phage ϕ80p lac carrying lac-operon (i +o+z+y+), the transcription of the lac-operon was studied in vitro. The lac-specific mRNA having a molecular size of 28S was detected by hybridizing messenger RNA (mRNA) from induced cells (diploid for the lac-operon) to the ϕ80p lac DNA. The hybridization of lac-specific mRNA to ϕ80p lac DNA was almost completely competed by RNA (ϕ80p lac RNA) synthesized in the RNA polymerase reaction primed by ϕ80p lac DNA. Therefore it was concluded that ϕ80p lac RNA synthesized in vitro includes mRNA corresponding to an almost entire region of the lac-operon.

It was clearly shown that ϕ80p lac RNA consisted of two kinds of RNA, one hybridizable to both ϕ80p lac DNA and ϕ80 DNA and the other (ΔRNA) only to ϕ80p lac DNA. The ΔRNA is assumed to be derived from genes incorporated from F′-13 into ϕ80p lac. The ΔRNA forms 30∼40% of the total ϕ80p lac RNA. 60% of the ΔRNA was shown to correspond to lac mRNA. The ΔRNA synthesis occured immediately after the reaction has started and the RNA size gew up to 30S by 10 min.

A part of ϕ80p lac RNA was also hybridizable to F′-lac RNA in contrast with ϕ80 RNA.

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Author notes

  1. Yoshiko Iida

    Present address: Institute of Physiology, Czechoslovak Academy of Sciences, Budějovická, 1083, Prague 4, Czechoslovakia

Authors and Affiliations

  1. The Cancer Research Institute, Kanazawa University, Kanazawa, Japan

    Yoshiko Iida & Tadanori Kameyama

  2. Faculty of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan

    Yasumi Ohshima & Tadao Horiuchi

Authors
  1. Yoshiko Iida
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  2. Tadanori Kameyama
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  3. Yasumi Ohshima
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  4. Tadao Horiuchi
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Iida, Y., Kameyama, T., Ohshima, Y. et al. Regulation of the lactose operon in vitro . Molec. Gen. Genetics 106, 296–306 (1970). https://doi.org/10.1007/BF00324047

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  • DOI: https://doi.org/10.1007/BF00324047

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