SU1203107A1 - Method of isolating growth factor for cultivated fruit fly cells - Google Patents
Method of isolating growth factor for cultivated fruit fly cells Download PDFInfo
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- SU1203107A1 SU1203107A1 SU843719812A SU3719812A SU1203107A1 SU 1203107 A1 SU1203107 A1 SU 1203107A1 SU 843719812 A SU843719812 A SU 843719812A SU 3719812 A SU3719812 A SU 3719812A SU 1203107 A1 SU1203107 A1 SU 1203107A1
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- SU
- USSR - Soviet Union
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- growth factor
- fruit fly
- cells
- activity
- fly cells
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- Peptides Or Proteins (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Description
Изобретение относитс к области биохимии, а именно к способам вьще- лени и очистки агента (фактора, стимулирующего рост культивируемых клеток насекомых.The invention relates to the field of biochemistry, namely to methods for the purification and purification of an agent (a factor that stimulates the growth of cultured insect cells.
Фактор роста может быть использован в фундаментальных исследовани х по клеточной биологии, а также в прикладных медицинских и вирусологических исследовани х, в которыхThe growth factor can be used in basic research in cell biology, as well as in applied medical and virological studies, in which
Состав раствора солей щелочньгх металлов, ммольThe composition of the solution of salts of alkali metals, mmol
Nad 69; КС1 21; Na%P0 3,3; NaHCOj 4,2 NaCl 69; KCl 21; NaH,PO 3,2Nad 69; KC1 21; Na% P0 3.3; NaHCOj 4.2 NaCl 69; KCl 21; NaH, PO 3.2
Примечание. + - активность фактора роста обнаруживаетс .Note. + - growth factor activity is detected.
- - активность фактора роста не обнаруживаетс .- - growth factor activity is not detected.
Экстракт центрифугируют в течение 20 мин при 4000xq. Вы влено, что при исходной концентрации клеток 1-2 10 /мл экстракт (0,03 мл/млThe extract is centrifuged for 20 minutes at 4000xq. It was revealed that with the initial cell concentration of 1-2 10 / ml extract (0.03 ml / ml
10-1510-15
Фактор роста, содержащийс в экстрактах , не удал етс диализом. Экстракт подвергают тепловой обработке в течение 5 мин дл удалени примесей при температурах, представленных в табл.3.The growth factor contained in the extracts is not removed by dialysis. The extract is heat treated for 5 minutes to remove impurities at the temperatures shown in Table 3.
ТаблицаЗTable3
Температура обработкиProcessing temperature
Вли ние на активность фактора ростаEffect on growth factor activity
60 С60 С
Не снижаетDoes not reduce
100 С100 C
Снижает в 5-6 разReduces 5-6 times
примен етс культура клеток насекомых .insect cell culture is used.
Цель изобретени - повышение активности целевого продукта. i) Пример. Провод т гомогенизацию куколок насекомых в гомогенизаторе Поттера в 3 мл раствора соле (.елочных металлов на 1 г куколок при 4 Ci Раствор солей щелочных ме- 10 таллов берут в соотношении, представленном в табл,1.The purpose of the invention is to increase the activity of the target product. i) Example. Conduct the homogenization of insect pupae in a Potter homogenizer in 3 ml of salt solution (alkaline metals per 1 g of pupae at 4 Ci. A solution of alkali metal salts is taken in the ratio shown in Table 1.
Таблица 1Table 1
Температура гомогенизацииHomogenization temperature
А- СA- C
среды необходим дл роста пересеваемой и первичной культуры клеток. Данные отражены :в табл.2,the medium is necessary for the growth of transplanted and primary cell culture. Data reflected: in table 2,
Таблица 2table 2
10-1510-15
Дл вы влени природы фактора роста экстракт обрабатьшают трипсином (традиционным способом) . Резкое снижение активности действующего начала, после обработки трипсином показало,To reveal the nature of the growth factor, the extract is treated with trypsin (in the traditional way). A sharp decrease in the activity of the active principle, after treatment with trypsin, showed
что фактор роста вл етс белком или содержит белковый компонент.that the growth factor is a protein or contains a protein component.
Молекул рный вес фактора роста составл ет 200 000 по результатам гельфильтрации на колонке с сефаро- зой 4В,The molecular weight of the growth factor is 200,000 by gel filtration on a Sepharose 4B column,
Дл очистки фактора роста примен ют хроматографию на ДЭАЭ-целлюлозеFor the purification of growth factor, chromatography on DEAE-cellulose is used.
51203107 451203107 4
(ДЕ-23, Whatman.). Активное началоПредлагаемое изобретение позвол - (DE-23, Whatman.). Active principle The proposed invention allows -
элюируетс 0,2 М NaCl. При меньшейет получить белковый фактор роста наконцентрации NaCl (0,1 М) активностьсёкомых, который интенсивно стимулив элюате не обнаруживаетс . В про-рует рост эмбриональных клеток в куль- цессе очистки активность фактора рос- 5 туре - количество клеток культуры eluted with 0.2 M NaCl. At a lesser level, to obtain a protein growth factor of concentration of NaCl (0.1 M), activity of the glasses, which is not intensely detected by the eluate. B will undergo the growth of embryonic cells in a purification culture; activity of the growth factor - 5 — the number of culture cells
та в расчете на мг белка увеличи-при добавлении фактора роста увеливаетс в 5-10 раз.чиваетс в 10-15 раз.that, per mg of protein, increases with the addition of a growth factor, it increases by 5-10 times shorter by 10-15 times.
Claims (1)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| SU843719812A SU1203107A1 (en) | 1984-03-30 | 1984-03-30 | Method of isolating growth factor for cultivated fruit fly cells |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| SU843719812A SU1203107A1 (en) | 1984-03-30 | 1984-03-30 | Method of isolating growth factor for cultivated fruit fly cells |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| SU1203107A1 true SU1203107A1 (en) | 1986-01-07 |
Family
ID=21110936
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| SU843719812A SU1203107A1 (en) | 1984-03-30 | 1984-03-30 | Method of isolating growth factor for cultivated fruit fly cells |
Country Status (1)
| Country | Link |
|---|---|
| SU (1) | SU1203107A1 (en) |
-
1984
- 1984-03-30 SU SU843719812A patent/SU1203107A1/en active
Non-Patent Citations (1)
| Title |
|---|
| Cholii W.J.et al. Purification of Drosophilla acidic ribosomal proteins.-Eur. I.Biochem., 1982, 127, № 1, pp.199-205. Wys3 C. at al. A cationic low molecular Weight growth factor from Drosophilla melanogaster and the nutritional requirements of KcHF cells-1 insect.-Biochem, 1982, V.12, № 5, pp.515-222. * |
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