SU946517A1 - Method of preparing complexes of dna with protein for electron-microscopic investigations - Google Patents

Method of preparing complexes of dna with protein for electron-microscopic investigations Download PDF

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Publication number
SU946517A1
SU946517A1 SU792837452A SU2837452A SU946517A1 SU 946517 A1 SU946517 A1 SU 946517A1 SU 792837452 A SU792837452 A SU 792837452A SU 2837452 A SU2837452 A SU 2837452A SU 946517 A1 SU946517 A1 SU 946517A1
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USSR - Soviet Union
Prior art keywords
dna
protein
electron
complexes
microscopic investigations
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SU792837452A
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Russian (ru)
Inventor
Юрий Юзефович Венгеров
Владимир Иванович Попенко
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Институт молекулярной биологии АН СССР
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Description

Изобретение ссгноситс  к медицине/ а именно к электронно-микроскопическим исследовани м биологических макромолекул и их взаимодействи .The invention is related to medicine / specifically to electron microscopic studies of biological macromolecules and their interaction.

Известен способ определени  комплексов ДНК с белком дл  электронномикроскопического исследовани  путем смешивани  растворов ДНК и белка, перенесени  их на пленки-подложки и контрастировани  1.A known method for determining DNA complexes with a protein for electron microscopic examination by mixing DNA solutions and protein, transferring them to substrate films and contrasting 1.

Однако известный способ не обеспечивает высокой точности, так как не позвол ет исследовать динамику структурных изменений, происход щих при присоединении белка к ДНК.However, the known method does not provide high accuracy, since it does not allow one to study the dynamics of structural changes that occur when a protein is attached to DNA.

Цель изобретени  - повышение точности способа, вы вление комплексов и изучение динамики их образовани .The purpose of the invention is to improve the accuracy of the method, detect the complexes and study the dynamics of their formation.

Поставленна  цель достигаетс  тем, что при осуществлении приготовлени  способа комплексов ДНК с белком дл  электронно-микроскопического исследовани  осуществл ют путем смешивани  растворов ДНК и белка, перенесени  их на пленки-подложки и контрастировани , раствор ДНК наслаи вают на поверхность гипофазы раствора белка.The goal is achieved by the fact that, when carrying out the preparation of the method of DNA complexes with protein for electron microscopic investigation, is performed by mixing DNA solutions and protein solutions, transferring them to substrate films and contrasting, the DNA solution is layered on the surface of the hypophase of the protein solution.

Пример. Исследование структуры комплексов ДНК с гистоном Hi,Example. Study of the structure of DNA complexes with histone Hi,

Каплю раствора ДНК (5 мгк/мл в 25 мМ ТЭА-нес буфере) осторожно наслаивают на поверхность гипофазы, содержащей 2 мкг/мл гистона Hi в присутствии 0,14 М NaCt.A drop of DNA solution (5 mg / ml in 25 mM TEA-carried buffer) is carefully layered on the surface of a hypophase containing 2 μg / ml of Hi histone in the presence of 0.14 M NaCt.

Образование пленки ДНК на поверисности контролируют по меткам талька . Затем через различные промежутки времени касаютс  полученного п т10 на электронно-микроскопической сеточкой , покрытой коллодиевой пленкой-подложкой . Избыток жидкости отсасывают фильтровальной бумагой. Сеточку контрастируют круговым напы15 лением сплавом Pt-Pd. Просмотр производ т на электронном микроскопе.The formation of DNA films on the surface is controlled by talcum labels. Then, at various intervals, the obtained pt10 is applied on an electron microscopic network coated with a collodium film substrate. Excess liquid is sucked off with filter paper. The mesh is contrasted by circular deposition of Pt – Pd alloy. View produced on an electron microscope.

Предлагаемый способ дает возможность исследовать динамику образовани  комплексов, изучать их струк20 туру, а также прослеживать изменени  конформации индивидуальных молекул ДНК при присоединении к ним молекул белка, наход щегос  в гипофазе , и расшир ет возможности элект25 ронно-микроскопического изучени  .биологических макромолекул.The proposed method makes it possible to investigate the dynamics of the formation of complexes, study their structure, as well as track the changes in the conformation of individual DNA molecules when protein molecules that are in the hypophase join them, and expands the possibilities of electronic microscopic study of biological macromolecules.

Claims (1)

Формула изобретени Invention Formula Способ приготовлени  комплексов 30 ДНК с белком дл  электронно-микро3 946517.4Method of preparing 30 DNA complexes with protein for electron micro 3 946517.4 скопического исследовани  путемна поверхность гипофазы раствора белсмешивани  растворов ДНК и белка,ка. . перенесени  их на пленки-подложкиScientifically study the hypophase surface of a solution of protein mixing of DNA and protein solutions, ka. . transfer them to the substrate film и контрастировани , отличаю-Источники информации,and contrasting, distinguish-Sources of information, щ и и с   тем, что, с целью повы-прин тые во внимание при экспертизеu and with the fact that, in order to be taken into account in the examination шени  точности способа/ вы влени 5 j. Cole R.D tawson C.M., Usiкомплексов и изучени  динамики ихang M.W. Cold Spring Harboy Sympoобразовани , раствор ДНК наслаивают slum Quantity Biology. 1977, p.253.Chance of accuracy of the method / manifestation 5 j. Cole R.D. tawson C.M., Usicomplexes and studying their dynamicsang M.W. Cold Spring Harboy Sympathy, DNA solution layers slum Quantity Biology. 1977, p.253.
SU792837452A 1979-06-29 1979-06-29 Method of preparing complexes of dna with protein for electron-microscopic investigations SU946517A1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
SU792837452A SU946517A1 (en) 1979-06-29 1979-06-29 Method of preparing complexes of dna with protein for electron-microscopic investigations

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Application Number Priority Date Filing Date Title
SU792837452A SU946517A1 (en) 1979-06-29 1979-06-29 Method of preparing complexes of dna with protein for electron-microscopic investigations

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SU946517A1 true SU946517A1 (en) 1982-07-30

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