Chloroplast «cryptic» promoter can be activated upon their transfer to plant nuclear genome
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 rpS...
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| Дата: | 1996 |
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Інститут молекулярної біології і генетики НАН України
1996
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| Назва журналу: | Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| Цитувати: | Chloroplast «cryptic» promoter can be activated upon their transfer to plant nuclear genome / N.V. Grgzelyak, A.P. Galkin, L.V. Gening, Т.V. Medvedeva, L.G. Lioshina, О.V. Bulko, K.G. Gasaryan // Биополимеры и клетка. — 1996. — Т. 12, № 6. — С. 87-93. — Бібліогр.: 28 назв. — англ. |
Репозитарії
Digital Library of Periodicals of National Academy of Sciences of Ukraine| _version_ | 1859630374207881216 |
|---|---|
| author | Grgzelyak, N.V. Galkin, A.P. Gening, L.V. Medvedeva, T.V. Lioshina, L.G. Bulko, O.V. Gasaryan, K.G. |
| author_facet | Grgzelyak, N.V. Galkin, A.P. Gening, L.V. Medvedeva, T.V. Lioshina, L.G. Bulko, O.V. Gasaryan, K.G. |
| citation_txt | Chloroplast «cryptic» promoter can be activated upon their transfer to plant nuclear genome / N.V. Grgzelyak, A.P. Galkin, L.V. Gening, Т.V. Medvedeva, L.G. Lioshina, О.V. Bulko, K.G. Gasaryan // Биополимеры и клетка. — 1996. — Т. 12, № 6. — С. 87-93. — Бібліогр.: 28 назв. — англ. |
| collection | DSpace DC |
| container_title | Биополимеры и клетка |
| description | 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.
Фрагменти рослинної ДНК клонували перед кодуючою послідовністю безпромоторного гена неомщинфосфотрансферази-ІІ і відбирали ті з них, що мали властивість ініціювати транскрипцію у бактеріальних клітинах. Блот-гібридизація і аналіз нуклеотидної послідовності показали, що один з клонованих фрагментів відповідає 3'-кінцевій частині хлоропластного гена рибосомного білка S12 ( p6S12). При введенні генної конструкції 5' -p6S12-XAT-nos-3' у протопласти тютюну за допомогою обробки ПЭГом у пробах виявлялася активність ферменту хлорамфеніколацетилтрансферази (ХАТ). Ці факти свідчать про те, що промотороподібні «криптичні» послідовності, які знаходяться у хлоропластному геномі, можуть бути активовані внаслідок їх штучного або природного переносу у ядерний геном.
Фрагменты растительной ДНК клонировали перед кодирующей последовательностью беспро- моторного гена неомицин-фосфотрансферазы-I I и отбирали те из них, которые имели способность инициировать транскрипцию в бактериальных клетках. Блот-гибридизация и анализ нуклеотидной последовательности показали, что один из клонированных фрагментов соответствует 3'-концевой части хлоропластного генарибосомного белка S12. При введении генной конструкции 5'-p6S 12-ХAT-nos-З' в протопласты табака с помощью обработки ПЭГом в пробах обнаруживалась активность фермента хлорамфениколацетилтрансферазы (ХАТ). Эти факты свидетельствуют о том, что промотороподобные «криптические» последовательности, которые находятся в хлоропластном геноме, могут быть активированы вследствие их искусственного или природного переноса в ядерный геном.
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| first_indexed | 2025-12-07T13:11:10Z |
| format | Article |
| fulltext |
ISSN 0233-7657. Биополимеры и клетка. 1996. Т. 12. № 6
Chloroplast «cryptic» promoter can be activated upon their
transfer to plant nuclear genome
N. V. Grgzelyak*, A. P. Galkin, L. V. Gening1, Т. V. Medvedeva,
L. G. Lioshina, О. V. Bulko, K. G. Gasaryan1
Institute of Bioorganic and Petroleum Chemistry, National Academy of Sciences of Ukraine
5 Murmanskaya str., Kiyv, 252660, Ukraine
'institute of Molecular Genetics, Russian Academy of Sciences
46 Kurchatov sq., Moscow, 123182, Russia
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 in transgenic plant nuclear genome. Transcription start points in the transgenic plant
were localized withintherpS12 DNA coding sequences. After5'-rpSJ2-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
Introduction. Evolutionary gene transfer from chloroplast to nuclear genomes is
a corner-stone of the now widely accepted endosymbiotic theory [1 ].
Many of the genes encoding proteins integral to plastid metabolism which
were originally encoded in the chloroplast genome are thought to have been
transfered to the nucleus during the course of plant evolution. Indeed, a number
of evident examples of intracellular gene «migration» from chloroplast to nuclei
have been reported [2—4 ].
As such movement of a gene has recurrently occured along plant evolution,
then the most of the plastid genes have homologous counterparts in the nucleus,
where they were found as short (< 1.0 kb) or long (several kb) sequences 15].
Evidently, these sequences in nuclear DNA don't differ markedly from those in
plastome DNA [6, 7].
The DNA sequences that move between different genomes of the cell are a
good tool to analyse mechanisms involved in this process and to understand the
consequences of such transfer. As gene transfer is a currently routine technique
for many plant species, it may be used for studies modelling some natural
mechanisms of the genetic flux.
In our previous papers plant DNA segments with transcription-promoting
activity were selected from a pool of random tobacco nuclear DNA fragments
[8 ]. One of the isolated DNA fragments cloned in pDNt23 plasmid was further
studied in detail. It was sequenced and transgenic plants containing npt-II gene
downstream this DNA segment were regenerated [9 ].
^Correspondence address.
© N. V GRGZELYAK, A P. GALKIN, L. V. GENING. Т. V MEDVEDEVA, L. G. LIOSHINA. О. V BULKO.
К G GASARYAN. 1996
87
GRGZELYAK N. V. ET AL.
In this paper we report that this DNA fragment represents the exon 2 of
the chloroplast gene for ribosomal protein S12 (rpS12) with flanking sequences
and present an evidence that when this fragment may occur to be located in
front of some structural gene, new patterns of the gene regulation could arise
due to the «cryptic» controlling elements upon transfer to the plant nuclear
genome.
Materials and Methods. Isolation and analysis of nucleic acids from
plants. Plant DNA was obtained from cell nuclei of green leaves [10].
Chloroplast DNA was purified by phenol-chlorophorm deproteinization [11].
Total RNA from plants was obtained by centrifugation through CsCl [12]. The
5 -ends of transcripts were mapped by SI nuclease technique [13].
Cloning and analysis of constructed plasmids. Restriction digests, ligations,
transformations, DNA labelling and plasmid preparation were done by standard
methods [14].
DNA sequence data were analysed in an IBM-PS AT computer using the
«DNA-STAR» and «PCGENE» software packages.
Protoplast isolation and DNA transfer. Leaf protoplasts of Nicotiana
tabacum cv. Petit Havana SRI were isolated from aseptically grown plants and
transformed as described previously [15].
The dividing protoplasts were allowed to develop for 3 weeks without
selection and then kanamycin was added to a concentration of 150 mg/1. After
approximately 6 weeks in culture individual colonies growing in the selective
medium were visible and were picked onto agar-solidified media containing
kanamycin. A total of 27 colonies were recovered from the transformed
protoplasts and 54 from the positive control. However, these colonies showed
marked differences in their growth on the solidified media, therefore two well
proliferating calli were cut into pieces and transfered to agar medium containing
2 mg/1 BAP, 0,2 mg/1 IAA, 2 % sucrose, 150 mg/1 kanamycin to stimulate
morphogenesis. Seven plants were regenerated and used in further studies.
In transient expression assays plasmid DNA was introduced into tobacco
protoplasts by the method of [16].
Enzyme assays. Assays for npt-II were performed by a modification of [17 ]
as described by [18]. CAT activity were determined according to [19]. Each
gene construction was assayed at least for 3 times.
Results and Discussion. Characterization of the pDNt23 clone. In order to
find out whether the cloned sequence represents tobacco DNA, labelled pDNt23
plasmid was hybridized to tobacco genomic and chloroplast DNA cut with the
restriction enzyme.
As the DNA insert of the pDNt23 hybridized to both ctDNA and nDNA,
we decided to compare its nucleotide sequence to the published ctDNA sequence
using the «DNA-STAR» computer search program. It was found that the 470
bp insert of pDNt23 had 100 % homology with the EcoRI-BgUI fragment of
the published tobacco chloroplast DNA sequence. It includes the exon 2
sequence of the gene for rpS12 and flanking sequences. Homology begins at a
nucleotided 100 478 and extends to a nucleotide of 100 948. Another homology
region extends from a 141 580 nucleotide up to nucleotide 142 050 [20].
Thus, the ctDNA contains two copies of this sequence cloned, in each
segments of the inverted repeat apiece.
NPT-II activity in transgenic plants. To test whether a cloned rpS12
fragment could initiate transcription in the regenerated plants and to prove that
the kanamycin resistance of plant is due to the expression of chimeric npt-II
gene the pDNt23 plasmid was introduced into tobacco protoplasts by a direct
gene transfer method [15] and transgenic plants were regenerated.
To determine whether the cloned tobacco DNA fragment could direct
non-tissue-specific expression or whether this expression is only tissue-specific,
88
CHLOROPLAST •CRYPTIC* PROMOTER
Fig. 1. Comparison of the npt-II activity
in different plant organs of the transgenic
plant: a — leaves; b — stem; с — roots of
transgenic plant carrying the pDNt23
plasmid; d — leaves of the transgenic
plant with the pLGV23neo plasmid [21]
npt-II activities were analysed in leaf, stem and root extracts of the transgenic
plant. Transgenic plant carrying the pLGV23neo plasmid served as a positive
control [21 ]. In the last case the npt-II activity was determined only in leaves
for the nos promoter is known to function constitutively in all the plant organs.
The results are presented in Fig. 1. As can be seen, the expression of the
chimeric npt-II gene product revealed highest activity in roots, followed by the
stem and near background activity in leaf tissue.
Mapping of the 5'-ends of transcripts synthesized in tobacco. The presence
of the npt-11 activity in the transgenic plant extracts indicates the production of
npt-II messenger in plant cells and can be explained either by transcriptional
read through or transcription initiation at the rpSI2 sequence. To distinguish
between these possibilities, we mapped the 5'-ends of the corresponding
transcripts synthesized in transgenic tobacco.
For hybridization with the total mRNA of the transgenic plant, containing
the chimeric npt-II gene we used the EcoRI-Bglll fragment labelled with 32P
at the 5 -end of the BgUI restriction site. After hybridization with plant mRNA
and SI nuclease treatment of the hybrid we found two fragments protected from
hydrolysis. The calculated length of the protected fragments locates the
transcription start points between 146 and 149 nucleotides upstream from the
BgUI restriction site (Fig. 2). The first nucleotides in both cases are the
guanines.
These results indicate that transcription start points of the corresponding
mRNAs in transgenic plant are localized within the rpS12 DNA fragment.
In order to find out whether the cloned fragment is transcribed in
Fig. 2. Mapping of the 5'-ends of transcripts in tobacco cells:
a — fragments, protected from SI nuclease hydrolysis by total
mRNA isolated from tobacco transgenic plant, carrying pDNt23
plasmid; b — control DNA ladder. Number on the right indicate
the lengths of the marker fragments
89
GRGZELYAK N. V. ET AL.
nontransgenic plants we also hybridized nick translated pDNt23 DNA with total
RNA isolated from tobacco plant. It is interesting that no hybridization patterns
were observed in this case.
Ttansient assay in tobacco protoplasts. It may also be assumed that
transcription initiation at rpS12 DNA fragment in transgenic plant can be
caused by position effect. In other words the rpS12 fragment alone could lack
the ability to initiate transcription and could be functionally active only if
flanked by some regulatory sequences.
To rule out this possibility we inserted rpS12 DNA fragment in front of
the CAT gene and tested its ability to initiate transcription by transient assay
in tobacco protoplasts. In this case the influence of nuclear rpS12 flanking
sequences is completely eliminated.
The presence of CAT activity in transformed tobacco protoplasts confirms
the ability of the cloned fragment to confer expression in plant cells (Fig. 3).
We have found that the previously isolated and characterized DNA
fragment from the pDNt23 plasmid represents the З'-part of the chloroplast
gene for rpSJ2. It consists of exon 2 of the rpS12 gene together with its flanking
sequences.
The rpS12 gene is known as a «divided» gene because its 5 -part is located
28 kbp downstream from the exon 2 in IRb on the same strand or 86 kbp
downstream the З'-part in IRa on the opposite strand [20, 22, 23].
Nevertheless, it is unknown whether this fact relates to the found ability of the
cloned fragment to initiate transcription.
Comparison of the cloned rpS12 DNA with sequences registered in the
EMBL DNA database revealed strong homology between analysed fragment
cloned in pDNt23 and corresponding rpS12 regions in chloroplasts of maize,
rice, soybean, livewort etc. Moreover, we also found a high degree (96 %) of
sequence homology between cloned rpSJ2 tobacco DNA and variable copy
number DNA sequences from the rice embryo genome [24]. Copy number of
this sequence changes during rice cell redifferentiation and growth and the
authors suggests that these and other sequences on the inverted repeat structure
of chloroplast DNA may have the character of a movable genetic element.
In view of this data, it may be reasonable to assume that the ability of
rpS12 DNA fragment to initiate transcription can bring in new gene regulation
patterns upon its integration in front of structural nuclear gene. Positive results
of such intracellular gene transfer have been reported [2 ].
Fig. 3. CAT activity in plant protoplasts transformed with pDNt23
plasmid: a — CAT activity in Escherichia coli celis, used as a marker
for acetylated forms of 14C-chloramphenicol; b — CAT activity in
tobacco protoplasts transformed with pDNt23 plasmid
14,
90
CHLOROPLAST «CRYPTIC» PROMOTER
Our experiments demostrate that insertion of this sequence in front of the
reporter npt-II gene lead to the organ-specific expression of the chimeric npt-II
gene. It is, of course, improbable, that the rpS12 fragment contains the
necessary ds-elements for tissue-specific expression. So, one can assume that
this may be due to the site of rpSJ2 DNA integration in the nuclear genome.
Unfortunately, we couldn't study this phenomenon more carefully because all
transgenic plants used in these experiments were probably originated from one
callus.
In order to verify whether the npt-II activity in transgenic plants is due to
the specific character of the cloned rpS12 DNA fragment we have mapped the
5'-ends of the mRNA transcripts in transgenic plant. The SI nuclease protection
data clearly indicate that transcription start points for the chimeric npt-II gene
are localized within the rpS12 DNA fragment.
An analysis of the rpS12 sequence shows that the nearest perfect eukaryotic
TATA-box is located at position (-115 bp) from the transcription start in plant
cells.
This is too far to be recognized by the RNA polymerase II transcription
complex, because in plants TATA-box is located at a average distance of
32+7 bp upstream from the transcription start points [25 ]. However, there are
a number of eukaryotic genes which do not contain TATA-boxes in the
promoters, but the regulation mechanisms of these genes are still unclear [26 ].
No final conclusions can therefore be drawn concerning the mechanism of
transcription initiation by the rpS12 DNA fragment.
The results described in this report indicate that although the rpS12 DNA
fragment is not a classical promoter, it can be recognized by eukaryotic RNA
polymerase. Recently, the same properties have been shown for plastid psbA
promoter of tobacco [27 ].
In conclusion, our results allow to suggest that «cryptic» regulatory
elements exists in chloroplast or perhaps in nuclear genome which can be
activated as a result of their translocation. Though in our experiments the role
of the natural genetic flux was modelled by gene transfer technique, we can
assume that natural DNA integration occures during artificial genetic
transformation [28 ].
The role of such translocation events in evolution and the frequency of the
occurance of «cryptic» controlling elements within plant genomes remain to be
determined.
H. В. Гржеляк, А. П. Галкін, Л. В. Генінг, Т. В. Медведева, Л. Г. Льоиіина, О. В. Булко, К. Г. Газарян
Активація хлоропластних «криптичних» промоторних послідовностей при переносі їх у
рослинний ядерний геном
Резюме
Фрагменти рослинної ДНК клонували перед кодуючою послідовністю безпромоторного гена нео-
мщинфосфотрансферази-ІІ і відбирали ті з них, що мали властивість ініціювати транскрипцію
у бактеріальних клітинах. Блот-гібридизація і аналіз нуклеотидної послідовності показали, що
один з клонованих фрагментів відповідає 3'-кінцевій частині хлоропластного гена рибосомного
білка S12 ( p6S12). При введенні генної конструкції 5' -p6S12-XAT-nos-3' у протопласти тютюну
за допомогою обробки ПЭГом у пробах виявлялася активність ферменту хлорамфеніколацетил-
трансферази (ХАТ). Ці факти свідчать про те, що промотороподібні «криптичні» послідовно-
сті, які знаходяться у хлоропластному геномі, можуть бути активовані внаслідок їх штучного
або природного переносу у ядерний геном.
91
GRGZELYAK N. V. ET AL.
H. В. Гржеляк, А. П. Галкин, Л. В. Генинг, Т. В. Медведева, Л. Г. Леиіина, О. В. Булко, К. Г. Газарян
Активация хлоропластных «криптических» промоторных последовательностей при переносе их
в растительный ядерный геном
Резюме
Фрагменты растительной ДНК клонировали перед кодирующей последовательностью беспро-
моторного гена неомицин-фосфотрансферазы-I I и отбирали те из них, которые имели способ-
ность инициировать транскрипцию в бактериальных клетках. Блот-гибридизация и анализ нук-
леотидной последовательности показали, что один из клонированных фрагментов соответст-
вует 3'-концевой части хлоропластного генарибосомного белка SJ2. При введении генной конст-
рукции 5'-p6S 12-ХAT-nos-З' в протопласты табака с помощью обработки ПЭГом в пробах обна-
руживалась активность фермента хлорамфениколацетилтрансферазы (ХАТ). Эти факты сви-
детельствуют о том, что промотороподобные «криптические» последовательности, которые
находятся в хлоропластном геноме, могут быть активированы вследствие их искусственного
или природного переноса в ядерный геном.
REFERENCES
1. Weeden N. F. Genetic and biochemical implications of the endosymbiotic origin of the
chloroplasts / / J. Мої. Evol.—1981.—17.—P. 133—139.
2. Pichersky E., Tankley S. D. Chloroplast DNA sequences integrated into an intron of a tomato
nuclear gene 11 Мої. and Gen. Genet.—1988.—215.—P. 65—68.
3. Baldauf S. L., Palmer J. D. Evolutionary transfer of the chloroplast tufA gene to the nucleus
/ / . Nature.—1990.—344.—P. 262—265.
4. Oliver J. L., Marin A., Martinez-Zapater J. M. Chloroplast genes transferred to the nuclear
plant genome have adjusted to nuclear base composition and codon usage / / Nucl. Acids
Res.—1990.—18.—P. 65—73.
5. Du Jardin P. Homologies to plastid DNA in the nuclear and mitochondrial genomes of potato
11 Theor. and Appl. Genet.—1990.—79.—P. 807—812.
6. Timmis J. N., Scott N. S. Sequence homology between spinach nuclear and chloroplast
genomes / / Nature.—1983.—305.—P. 65—67.
7. Cheung W. J., Scott N. S. A continuous sequence in spinach nuclear DNA is homologous to
three separated sequences in chloroplast DNA / / Theor. and Appl. Genet.—1989.—77.—
P. 625—633.
8. Domansky N. N., Gening L. V., Galkin A. P. et al. Cloning of the Nicotiana tabacum nuclear
DNA sequences that work like promoters in Escherichia coli cells / / Докл. Акад. наук
СССР—1986.—291.—С. 1004—1008.
9. Domansky N. N., Gening L. V., Kovalenko P. G. et al. Cloning of the DNA fragment that has
promoter properties in the transgenic plants / / Молекуляр. биология.—1989.—23.—
С. 1391 — 1399.
10. Lichtenstein С. P., Draper J. Genetic engineering of plants DNA cloning / Ed. D. M. Glover.—
Oxford: IRL press, 1986.—Vol. 11.—P. 107.
11. Bookjons G.f Stumman В. M., Henningsen K. W. Preparation of chloroplast DNA from pea
plastids isolated in a medium of high ionic strength / / Anal. Biochem.—1984.—141.—P. 244—
247.
12. Chirgwin J. M., Przybyla A. E., McDonald R. J. et al. Isolation of biologically active ribonucleic
acids from sources enriched in ribonuclease / / Biochemistry.—1979.—218.—P. 5294—5299.
13. DiRita V. G., Gelwin S. B. Deletion analysis of the mannopine, synthase gene promoter in
sunflower crown gall tumors and Agrobacterium tumefasience 11 Мої. and Gen. Genet.—
1987.—207.—P. 233—241.
14. Maniatis Т., Fritsch E. E., Sambrook J. Molecular cloning: a laboratory manual.— New York:
Cold Spring Harbor Lab., 1987.—352 p.
15. Shillito R. D., Saul M. W., Paszkowski J. et al. High efficiency direct gene transfer to plants
/ / Bio/Technology.—1985.—3.—P. 1099—1103.
16. Prols M., Topfer R., Shell J., Steinbiss H.-H. Transient gene expression in tobacco protoplasts.
1. Time course of CAT appearence 11 Plant. Cell. Rep.—1988.—7.—P. 221—224.
17. Reiss В., Sprengler R., Will H., Schaller H. A new sensitive method for qualitative and
quantitative assay of neomycin phosphotransferase in crude cell extracts / / Gene.—1984.—
30.—P. 211—218.
18. Schreier P. H., Sefton E. A., Schell J., Bohnert H. J. The use of nuclear encoded sequences
to direct the light-regulated synthesis and transport of foreign protein into plant chloroplasts / /
EMBO J.—1985.—4.—P. 25—32.
92
CHLOROPLAST «CRYPTIC» PROMOTER
19. Odell J. Т., Knowlton S., Lin W., Mauvais C. J. Properties of an isolated transcription
stimulating sequence derived from the cauliflower mosaic virus 35S promoter 11 Plant. Мої.
Biol.—1988.—10.—P. 263—272.
20. Shinozaki K.t Ohme M., Tanaka M. et al. The complete nucleotide sequence of the tobacco
chloroplast genome: its gene organization and expression / / EMBO J.—1986.—2.—P. 987—
995.
21. Herrera-Estrella L.t De Block M., Messens E. et al. Chimeric genes as dominant selectable
markers in plant cells / / Ibid.—1983.—2.—P. 987—995.
22. Torazawa K., Hayashida NObokata J. et al. The 5'-part of the gene for ribosomal protein
S12 is located 30 kbp downstream from its З'-part in tobacco chloroplast genome / / Nucl. Acids
Res. —1986.—14.— P. 3143.
23. Zaita N., Torazawa КShinozaki K.f Sugiura M. Trans splicing in vivo: joining of transcripts
from the «divided» gene for ribosomal protein SJ2 in the chloroplasts of tobacco / / FEBS
Let.—1987.—210.—P. 153—156.
24. Kikuchi S.f Takaiva F.t Oono K. Variable copy number DNA sequences in rice / / Мої. and
Gen. Genet.—1987.—210.—P. 373—380.
25. Messing J., Geraghty D.t Heidecker G. et al. Plant gene structure / / Eds T. Kosuge, C. P.
Meredity, A. Hollaender. — New York: Plenum press, 1983.—P. 211—227.
26. Dynan W. S. Promoters for housekeeping genes 11 Trends Genet.—1986.—4.—P. 196—197.
27. Cornelissen M., Vandewiele M. Nuclear transcription activity of the tobacco plastid psbA
promoter / / Nucl. Acids Res.—1989.—17.—P. 19—28.
28. Pichersky E., Logsdon J. M., Jr., McGrath J. M.t Stasys R. A. Fragments of plastid DNA in
the nuclear genome of tomato: prevalence, chromosomal location, and possible mechanism of
integration / / Мої. and Gen. Genet.—1991 .—225.—P. 453—458.
УДК 577 21 4.625 Received 23.04.96
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| id | nasplib_isofts_kiev_ua-123456789-154285 |
| institution | Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| issn | 0233-7657 |
| language | English |
| last_indexed | 2025-12-07T13:11:10Z |
| publishDate | 1996 |
| publisher | Інститут молекулярної біології і генетики НАН України |
| record_format | dspace |
| spelling | Grgzelyak, N.V. Galkin, A.P. Gening, L.V. Medvedeva, T.V. Lioshina, L.G. Bulko, O.V. Gasaryan, K.G. 2019-06-15T12:21:29Z 2019-06-15T12:21:29Z 1996 Chloroplast «cryptic» promoter can be activated upon their transfer to plant nuclear genome / N.V. Grgzelyak, A.P. Galkin, L.V. Gening, Т.V. Medvedeva, L.G. Lioshina, О.V. Bulko, K.G. Gasaryan // Биополимеры и клетка. — 1996. — Т. 12, № 6. — С. 87-93. — Бібліогр.: 28 назв. — англ. 0233-7657 DOI:http://dx.doi.org/10.7124/bc.00045A https://nasplib.isofts.kiev.ua/handle/123456789/154285 577.214.625 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. Фрагменти рослинної ДНК клонували перед кодуючою послідовністю безпромоторного гена неомщинфосфотрансферази-ІІ і відбирали ті з них, що мали властивість ініціювати транскрипцію у бактеріальних клітинах. Блот-гібридизація і аналіз нуклеотидної послідовності показали, що один з клонованих фрагментів відповідає 3'-кінцевій частині хлоропластного гена рибосомного білка S12 ( p6S12). При введенні генної конструкції 5' -p6S12-XAT-nos-3' у протопласти тютюну за допомогою обробки ПЭГом у пробах виявлялася активність ферменту хлорамфеніколацетилтрансферази (ХАТ). Ці факти свідчать про те, що промотороподібні «криптичні» послідовності, які знаходяться у хлоропластному геномі, можуть бути активовані внаслідок їх штучного або природного переносу у ядерний геном. Фрагменты растительной ДНК клонировали перед кодирующей последовательностью беспро- моторного гена неомицин-фосфотрансферазы-I I и отбирали те из них, которые имели способность инициировать транскрипцию в бактериальных клетках. Блот-гибридизация и анализ нуклеотидной последовательности показали, что один из клонированных фрагментов соответствует 3'-концевой части хлоропластного генарибосомного белка S12. При введении генной конструкции 5'-p6S 12-ХAT-nos-З' в протопласты табака с помощью обработки ПЭГом в пробах обнаруживалась активность фермента хлорамфениколацетилтрансферазы (ХАТ). Эти факты свидетельствуют о том, что промотороподобные «криптические» последовательности, которые находятся в хлоропластном геноме, могут быть активированы вследствие их искусственного или природного переноса в ядерный геном. en Інститут молекулярної біології і генетики НАН України Биополимеры и клетка Chloroplast «cryptic» promoter can be activated upon their transfer to plant nuclear genome Активація хлоропластних «криптичних» промоторних послідовностей при переносі їх у рослинний ядерний геном Активация хлоропластных «криптических» промоторных последовательностей при переносе их в растительный ядерный геном Article published earlier |
| spellingShingle | Chloroplast «cryptic» promoter can be activated upon their transfer to plant nuclear genome Grgzelyak, N.V. Galkin, A.P. Gening, L.V. Medvedeva, T.V. Lioshina, L.G. Bulko, O.V. Gasaryan, K.G. |
| title | Chloroplast «cryptic» promoter can be activated upon their transfer to plant nuclear genome |
| title_alt | Активація хлоропластних «криптичних» промоторних послідовностей при переносі їх у рослинний ядерний геном Активация хлоропластных «криптических» промоторных последовательностей при переносе их в растительный ядерный геном |
| title_full | Chloroplast «cryptic» promoter can be activated upon their transfer to plant nuclear genome |
| title_fullStr | Chloroplast «cryptic» promoter can be activated upon their transfer to plant nuclear genome |
| title_full_unstemmed | Chloroplast «cryptic» promoter can be activated upon their transfer to plant nuclear genome |
| title_short | Chloroplast «cryptic» promoter can be activated upon their transfer to plant nuclear genome |
| title_sort | chloroplast «cryptic» promoter can be activated upon their transfer to plant nuclear genome |
| url | https://nasplib.isofts.kiev.ua/handle/123456789/154285 |
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