Biopolym. Cell. 1996; 12(6):87-93.
Chloroplast «cryptic» promoter can be activated upon their transfer to plant nuclear genome
1Grgzelyak N. V., 1Galkin A. P., 2Gening L. V., 1Medvedeva T. V., 1Lioshina L. G., 1Bulko O. V., 2Gasaryan K. G.
  1. Institute of Bioorganic Chemistry and Petrochemistry, NAS of Ukraine
    1, Murmans'ka Str., Kyiv, Ukraine, 02094
  2. Institute of Molecular Genetics, Academy of Sciences of the USSR
    2, Academician Kurchatov Sq., Moscow, Russian Federation, 123182


It is shown that a DNA fragment containing the previously described exon 2 sequence of the chloroplast gene for ribosomal protein S12 can determine expression of the reporter npt-II gene intransgenic plant nuclear genome. Transcription start points inthetransgenic plant were localized within the rpS12 DNA coding sequences. After 5'-rpS12-CAT-nos-3' gene construction has been introduced to tobacco protoplasts by PEG-treatment, chloramphenicol acetyltransferase (CAT) enzyme activity was detectable by transient assays. These facts indicate that «cryptic» promoter-like sequences exist in chloroplast genome which can be activated as a result of their artifical or natural transfer to the nuclei.


[1] Weeden NF. Genetic and biochemical implications of the endosymbiotic origin of the chloroplast. J Mol Evol. 1981;17(3):133-9.
[2] Pichersky E, Tanksley SD. Chloroplast DNA sequences integrated into an intron of a tomato nuclear gene. Mol Gen Genet. 1988;215(1):65-8.
[3] Baldauf SL, Palmer JD. Evolutionary transfer of the chloroplast tufA gene to the nucleus. Nature. 1990;344(6263):262-5.
[4] Oliver JL, Mar?n A, Mart?nez-Zapater JM. Chloroplast genes transferred to the nuclear plant genome have adjusted to nuclear base composition and codon usage. Nucleic Acids Res. 1990;18(1):65-73.
[5] du Jardin P. Homologies to plastid DNA in the nuclear and mitochondrial genomes of potato. Theor Appl Genet. 1990;79(6):807-12.
[6] Timmis JN, Scott NS. Sequence homology between spinach nuclear and chloroplast genomes. Nature. 1983;305(5929):65–7.
[7] Cheung WY, Scott NS. A contiguous sequence in spinach nuclear DNA is homologous to three separated sequences in chloroplast DNA. Theor Appl Genet. 1989;77(5):625-33.
[8] Domansky NN, Gening LV, Galkin AP et al. Cloning of the Nicotiana tabacum nuclear DNA sequences that work like promoters in Escherichia coli cells. Dokl Akad Nauk SSSR; 1986; 291: 1004-8.
[9] Domanski? NN, Gening LV, Kovalenko PG, Medvedeva TV, Galkin AP, Gazarian KG. [Cloning of tobacco DNA fragment with promoter properties in a transgenic plant]. Mol Biol (Mosk). 1989;23(5):1391-9.
[10] Lichtenstein CP, Draper J. Genetic engineering of plants DNA cloning. Ed. D. M. Glover. Oxford: IRL press, 1986. Vol. 11: 107.
[11] Bookjans G, Stummann BM, Henningsen KW. Preparation of chloroplast DNA from pea plastids isolated in a medium of high ionic strength. Anal Biochem. 1984;141(1):244-7.
[12] Chirgwin JM, Przybyla AE, MacDonald RJ, Rutter WJ. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979;18(24):5294-9.
[13] DiRita VJ, Gelvin SB. Deletion analysis of the mannopine synthase gene promoter in sunflower crown gall tumors and Agrobacterium tumefaciens. Mol Gen Genet. 1987;207(2-3):233-41.
[14] Maniatis T, Fritsch EE, Sambrook J. Molecular cloning: a laboratory manual. New York: Cold Spring Harbor Lab., 1987. 352 p.
[15] Shillito RD, Saul MW, Paszkowski J, M?ller M, Potrykus I. High Efficiency Direct Gene Transfer to Plants. Bio/Technology. 1985;3(12):1099–103.
[16] Pr?ls M, T?pfer R, Schell J, Steinbi? HH. Transient gene expression in tobacco protoplasts: I. Time course of CAT appearance. Plant Cell Rep. 1988;7(4):221-4.
[17] Reiss B, Sprengel R, Will H, Schaller H. A new sensitive method for qualitative and quantitative assay of neomycin phosphotransferase in crude cell extracts. Gene. 1984;30(1-3):211-7.
[18] Schreier PH, Seftor EA, Schell J, Bohnert HJ. The use of nuclear-encoded sequences to direct the light-regulated synthesis and transport of a foreign protein into plant chloroplasts. EMBO J. 1985;4(1):25-32.
[19] Odell JT, Knowlton S, Lin W, Mauvais CJ. Properties of an isolated transcription stimulating sequence derived from the cauliflower mosaic virus 35S promoter. Plant Mol Biol. 1988;10(3):263-72.
[20] Shinozaki K, Ohme M, Tanaka M, Wakasugi T, Hayashida N, Matsubayashi T, Zaita N, Chunwongse J, Obokata J, Yamaguchi-Shinozaki K, Ohto C, Torazawa K, Meng BY, Sugita M, Deno H, Kamogashira T, Yamada K, Kusuda J, Takaiwa F, Kato A, Tohdoh N, Shimada H, Sugiura M. The complete nucleotide sequence of the tobacco chloroplast genome: its gene organization and expression. EMBO J. 1986;5(9):2043-2049.
[21] Herrera-Estrella L, Block MD, Messens E, Hernalsteens JP, Montagu MV, Schell J. Chimeric genes as dominant selectable markers in plant cells. EMBO J. 1983;2(6):987-95.
[22] Torazawa K, Hayashida N, Obokata J, Shinozaki K, Sugiura M. The 5' part of the gene for ribosomal protein S12 is located 30 kbp downstream from its 3' part in tobacco chloroplast genome. Nucleic Acids Res. 1986;14(7):3143.
[23] Zaita N, Torazawa K, Shinozaki K, Sugiura M. Trans splicing in vivo: joining of transcripts from the “divided” gene for ribosomal protein S12 in the chloroplasts of tobacco. FEBS Letters. 1987;210(2):153–6.
[24] Kikuchi S, Takaiwa F, Oono K. Variable copy number DNA sequences in rice. Mol Gen Genet. 1987;210(3):373-80.
[25] Messing J, Geraghty D, Heidecker G, Hu N-T, Kridl J, Rubenstein I. Plant gene structure. Eds T. Kosuge, C. P. Meredity, A. Hollaender. New York: Plenum press, 1983: 211-27.
[26] Dynan WS. Promoters for housekeeping genes. Trends Genet. 1986;2:196–7.
[27] Cornelissen M, Vandewiele M. Nuclear transcriptional activity of the tobacco plastid psbA promoter. Nucleic Acids Res. 1989;17(1):19-29.
[28] Pichersky E, Logsdon JM Jr, McGrath JM, Stasys RA. Fragments of plastid DNA in the nuclear genome of tomato: prevalence, chromosomal location, and possible mechanism of integration. Mol Gen Genet. 1991;225(3):453-8.