ΠΠΌΠΏΠΎΡΡ ΡΠ ΠΠ Π² ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠΈ Π΄ΡΠΎΠΆΠΆΠ΅ΠΉ: ΡΠΎΠ»Ρ ΠΏΡΠ΅Π΄ΡΠ΅ΡΡΠ²Π΅Π½Π½ΠΈΠΊΠ° ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΠ½ΠΎΠΉ Π»ΠΈΠ·ΠΈΠ»-ΡΠ ΠΠ-ΡΠΈΠ½ΡΠ΅ΡΠ°Π·Ρ ΠΈ ΡΡΠ½ΠΊΡΠΈΡ ΠΈΠΌΠΏΠΎΡΡΠΈΡΡΠ΅ΠΌΠΎΠΉ ΡΠ ΠΠ Π² ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΠ½ΠΎΠΌ ΠΌΠ°ΡΡΠΈΠΊΡΠ΅
ΠΠ²ΡΠΎΡ ΡΡΠΈΡΠ°Π΅Ρ ΡΠ²ΠΎΠΈΠΌ Π΄ΠΎΠ»Π³ΠΎΠΌ Π²ΡΡΠ°Π·ΠΈΡΡ Π³Π»ΡΠ±ΠΎΡΠ°ΠΉΡΡΡ ΠΏΡΠΈΠ·Π½Π°ΡΠ΅Π»ΡΠ½ΠΎΡΡΡ ΠΈ ΠΎΠ³ΡΠΎΠΌΠ½ΡΡ Π±Π»Π°Π³ΠΎΠ΄Π°ΡΠ½ΠΎΡΡΡ ΡΠ²ΠΎΠΈΠΌ Π½Π°ΡΡΠ½ΡΠΌ ΡΡΠΊΠΎΠ²ΠΎΠ΄ΠΈΡΠ΅Π»ΡΠΌ, ΠΠ³ΠΎΡΡ ΠΠ»Π΅ΠΊΡΠ°Π½Π΄ΡΠΎΠ²ΠΈΡΡ ΠΡΠ°ΡΠ΅Π½ΠΈΠ½Π½ΠΈΠΊΠΎΠ²Ρ ΠΈ ΠΠ²Π°Π½Ρ Π’Π°ΡΠ°ΡΠΎΠ²Ρ. ΠΠ½ΠΈ Π²ΡΠ΅ΠΌΠ΅ΡΠ½ΠΎ ΠΏΠΎΠΌΠΎΠ³Π°Π»ΠΈ ΠΌΠ½Π΅ ΠΏΡΠΈ ΠΏΠΎΡΡΠ°Π½ΠΎΠ²ΠΊΠ΅ ΡΠΊΡΠΏΠ΅ΡΠΈΠΌΠ΅Π½ΡΠΎΠ², ΠΈΠ½ΡΠ΅ΡΠΏΡΠ΅ΡΠ°ΡΠΈΠΈ ΡΠ΅Π·ΡΠ»ΡΡΠ°ΡΠΎΠ² ΠΈ Π½Π°ΠΏΠΈΡΠ°Π½ΠΈΠΈ ΡΡΠ°ΡΠ΅ΠΉ ΠΈ Π΄ΠΈΡΡΠ΅ΡΡΠ°ΡΠΈΠΈ. ΠΠΎΠ»Π΅Π΅ ΡΠΎΠ³ΠΎ, ΠΏΠΎ ΠΏΡΠΎΡΠ΅ΡΡΠ²ΠΈΠΈ ΡΠ΅ΠΌΠΈ Π»Π΅Ρ ΡΠΎΠ²ΠΌΠ΅ΡΡΠ½ΠΎΠΉ ΡΠ°Π±ΠΎΡΡ Ρ ΡΠΌΠ΅Π»ΠΎ ΠΌΠΎΠ³Ρ Π½Π°Π·Π²Π°ΡΡ ΠΈΡ ΡΠ²ΠΎΠΈΠΌΠΈ Π΄ΡΡΠ·ΡΡΠΌΠΈ, Ρ ΠΊΠΎΡΠΎΡΡΠΌΠΈ ΠΌΠ½Π΅… Π§ΠΈΡΠ°ΡΡ Π΅ΡΡ >
- Π‘ΠΎΠ΄Π΅ΡΠΆΠ°Π½ΠΈΠ΅
- ΠΡΠ΄Π΅ΡΠΆΠΊΠ°
- ΠΠΈΡΠ΅ΡΠ°ΡΡΡΠ°
- ΠΡΡΠ³ΠΈΠ΅ ΡΠ°Π±ΠΎΡΡ
- ΠΠΎΠΌΠΎΡΡ Π² Π½Π°ΠΏΠΈΡΠ°Π½ΠΈΠΈ
Π‘ΠΎΠ΄Π΅ΡΠΆΠ°Π½ΠΈΠ΅
- Π‘ΠΏΠΈΡΠΎΠΊ ΠΈΡΠΏΠΎΠ»ΡΠ·ΡΠ΅ΠΌΡΡ ΡΠΎΠΊΡΠ°ΡΠ΅Π½ΠΈΠΉ
- 1. ΠΠ²Π΅Π΄Π΅Π½ΠΈΠ΅
- 2. ΠΠ±Π·ΠΎΡ Π»ΠΈΡΠ΅ΡΠ°ΡΡΡΡ. ΠΠΌΠΏΠΎΡΡ ΠΌΠ°ΠΊΡΠΎΠΌΠΎΠ»Π΅ΠΊΡΠ» Π² ΠΌΠΈΡΠΎΡ
ΠΎΠ½Π΄ΡΠΈΠΈ
- 2. 1. ΠΠΌΠΏΠΎΡΡ Π±Π΅Π»ΠΊΠΎΠ² Π² ΠΌΠΈΡΠΎΡ
ΠΎΠ½Π΄ΡΠΈΠΈ
- 2. 1. 1. Π‘ΠΈΠ³Π½Π°Π»ΡΠ½ΡΠ΅ ΠΏΠΎΡΠ»Π΅Π΄ΠΎΠ²Π°ΡΠ΅Π»ΡΠ½ΠΎΡΡΠΈ ΠΈΠΌΠΏΠΎΡΡΠΈΡΡΠ΅ΠΌΡΡ Π±Π΅Π»ΠΊΠΎΠ²
- 2. 1. 2. ΠΡΠ΅ΡΡΠ°Π½ΡΠ»ΠΎΠΊΠ°ΡΠΈΠΎΠ½Π½ΠΎΠ΅ ΡΠ°Π·Π²ΠΎΡΠ°ΡΠΈΠ²Π°Π½ΠΈΠ΅ ΠΏΡΠ΅Π΄ΡΠ΅ΡΡΠ²Π΅Π½Π½ΠΈΠΊΠΎΠ² ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠ°Π»Ρ-Π½ΡΡ Π±Π΅Π»ΠΊΠΎΠ² ΠΈ ΠΈΡ ΡΡΠ°Π½ΡΠΏΠΎΡΡ ΠΊ ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΡΠΉ ΠΏΠΎΠ²Π΅ΡΡ Π½ΠΎΡΡΠΈ
- 2. 1. 3. Π’ΡΠ°Π½ΡΠ»ΠΎΠΊΠ°ΡΠΈΠΎΠ½Π½ΡΠΉ ΠΊΠΎΠΌΠΏΠ»Π΅ΠΊΡ Π²Π½Π΅ΡΠ½Π΅ΠΉ ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΡΠΉ ΠΌΠ΅ΠΌΠ±ΡΠ°Π½Ρ (Π’ΠΠ)
- 2. 1. 4. ΠΡΡΡΠ°ΠΈΠ²Π°Π½ΠΈΠ΅ Π±Π΅Π»ΠΊΠΎΠ² Π²ΠΎ Π²Π½Π΅ΡΠ½ΡΡ ΠΌΠ΅ΠΌΠ±ΡΠ°Π½Ρ ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠΉ ΠΏΡΠΈ ΠΏΠΎΠΌΠΎΡΠΈ ΠΠΠ-ΠΊΠΎΠΌΠΏΠ»Π΅ΠΊΡΠ°
- 2. 1. 5. ΠΠΌΠΏΠΎΡΡ Π±Π΅Π»ΠΊΠΎΠ² Π² ΠΌΠ΅ΠΆΠΌΠ΅ΠΌΠ±ΡΠ°Π½Π½ΠΎΠ΅ ΠΏΡΠΎΡΡΡΠ°Π½ΡΡΠ²ΠΎ ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠΉ
- 2. 1. 6. Π’ΡΠ°Π½ΡΠ»ΠΎΠΊΠ°ΡΠΈΠΎΠ½Π½ΡΠΉ ΠΊΠΎΠΌΠΏΠ»Π΅ΠΊΡ Π²Π½ΡΡΡΠ΅Π½Π½Π΅ΠΉ ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΡΠΉ ΠΌΠ΅ΠΌΠ±ΡΠ°Π½Ρ Π’1Π23 ΠΈ ΠΌΠΎΡΠΎΡ ΡΡΠ°Π½ΡΠ»ΠΎΠΊΠ°ΡΠΈΠΈ
- 2. 1. 7. Π’ΡΠ°Π½ΡΠ»ΠΎΠΊΠ°ΡΠΈΠΎΠ½Π½ΡΠΉ ΠΊΠΎΠΌΠΏΠ»Π΅ΠΊΡ Π²Π½ΡΡΡΠ΅Π½Π½Π΅ΠΉ ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΡΠΉ ΠΌΠ΅ΠΌΠ±ΡΠ°Π½Ρ Π’1Π22 ΠΈ ΠΌΠ°Π»ΡΠ΅ Π’Ρ-Π±Π΅Π»ΠΊΠΈ ΠΌΠ΅ΠΆΠΌΠ΅ΠΌΠ±ΡΠ°Π½Π½ΠΎΠ³ΠΎ ΠΏΡΠΎΡΡΡΠ°Π½ΡΡΠ²Π°
- 2. 1. 8. ΠΠΊΡΠΏΠΎΡΡ Π±Π΅Π»ΠΊΠΎΠ² ΠΈΠ· ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΠ½ΠΎΠ³ΠΎ ΠΌΠ°ΡΡΠΈΠΊΡΠ°
- 2. 2. ΠΠΌΠΏΠΎΡΡ Π ΠΠ Π² ΠΌΠΈΡΠΎΡ
ΠΎΠ½Π΄ΡΠΈΠΈ
- 2. 2. 1. ΠΠΌΠΏΠΎΡΡ ΡΠ ΠΠ Π² ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠΈ ΠΏΡΠΎΡΡΠ΅ΠΉΡΠΈΡ
- 2. 2. 2. ΠΠΌΠΏΠΎΡΡ ΡΠ ΠΠ Π² ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠΈ ΡΠ°ΡΡΠ΅Π½ΠΈΠΉ
- 2. 2. 3. ΠΠΌΠΏΠΎΡΡ ΡΠ ΠΠ Π² ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠΈ Π΄ΡΠΎΠΆΠΆΠ΅ΠΉ
- 2. 2. 4. ΠΠΌΠΏΠΎΡΡ Π ΠΠ Π² ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠΈ ΠΌΠ»Π΅ΠΊΠΎΠΏΠΈΡΠ°ΡΡΠΈΡ
- 2. 3. ΠΠΌΠΏΠΎΡΡ ΠΠΠ Π² ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠΈ
- 2. 1. ΠΠΌΠΏΠΎΡΡ Π±Π΅Π»ΠΊΠΎΠ² Π² ΠΌΠΈΡΠΎΡ
ΠΎΠ½Π΄ΡΠΈΠΈ
- 3. ΠΠ°ΡΠ΅ΡΠΈΠ°Π»Ρ ΠΈ ΠΌΠ΅ΡΠΎΠ΄Ρ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΡ
- 3. 1. ΠΠ°ΡΠ΅ΡΠΈΠ°Π»Ρ
- 3. 1. 1. Π Π΅Π°ΠΊΡΠΈΠ²Ρ
- 3. 1. 2. ΠΠΎΠΌΠΌΠ΅ΡΡΠ΅ΡΠΊΠΈΠ΅ Π½Π°Π±ΠΎΡΡ
- 3. 1. 3. ΠΠ½ΡΠΈΡΠ΅Π»Π°
- 3. 1. 4. ΠΡΠΈΠ±ΠΎΡΡ ΠΈ ΠΎΠ±ΠΎΡΡΠ΄ΠΎΠ²Π°Π½ΠΈΠ΅
- 3. 1. 5. Π¨ΡΠ°ΠΌΠΌΡ ΠΌΠΈΠΊΡΠΎΠΎΡΠ³Π°Π½ΠΈΠ·ΠΌΠΎΠ²
- 3. 1. 6. ΠΠΈΡΠ°ΡΠ΅Π»ΡΠ½ΡΠ΅ ΡΡΠ΅Π΄Ρ
- 3. 1. 7. ΠΠ΅Π½Π½ΠΎ-ΠΈΠ½ΠΆΠ΅Π½Π΅ΡΠ½ΡΠ΅ ΠΊΠΎΠ½ΡΡΡΡΠΊΡΠΈΠΈ
- 3. 1. 8. ΠΠ»ΠΈΠ³ΠΎΠ½ΡΠΊΠ»Π΅ΠΎΡΠΈΠ΄Ρ
- 3. 2. ΠΠ΅ΡΠΎΠ΄Ρ
- 3. 2. 1. ΠΠΎΠ»ΠΈΠΌΠ΅ΡΠ°Π·Π½Π°Ρ ΡΠ΅ΠΏΠ½Π°Ρ ΡΠ΅Π°ΠΊΡΠΈΡ (ΠΠ¦Π )
- 3. 2. 2. Π Π°ΡΡΠ΅ΠΏΠ»Π΅Π½ΠΈΠ΅ ΠΠΠ ΡΠ½Π΄ΠΎΠ½ΡΠΊΠ»Π΅Π°Π·Π°ΠΌΠΈ ΡΠ΅ΡΡΡΠΈΠΊΡΠΈΠΈ ΠΈ Π΄Π΅ΡΠΎΡΡΠΎ-ΡΠΈΠ»ΠΈΡΠΎΠ²Π°Π½ΠΈΠ΅
- 3. 2. 3. ΠΠΈΠ³ΠΈΡΠΎΠ²Π°Π½ΠΈΠ΅
- 3. 2. 4. ΠΠΏΡΠ΅Π΄Π΅Π»Π΅Π½ΠΈΠ΅ Π½ΡΠΊΠ»Π΅ΠΎΡΠΈΠ΄Π½ΠΎΠΉ ΠΏΠΎΡΠ»Π΅Π΄ΠΎΠ²Π°ΡΠ΅Π»ΡΠ½ΠΎΡΡΠΈ ΠΠΠ ΠΌΠ΅ΡΠΎΠ΄ΠΎΠΌ Π‘ΡΠ½Π³Π΅ΡΠ°
- 3. 2. 5. Π’ΡΠ°Π½ΡΡΠΎΡΠΌΠ°ΡΠΈΡ ΠΊΠ»Π΅ΡΠΎΠΊ E. col
- 3. 2. 6. Π’ΡΠ°Π½ΡΡΠΎΡΠΌΠ°ΡΠΈΡ ΠΊΠ»Π΅ΡΠΎΠΊ Saccharomyces cerevisiae
- 3. 2. 7. ΠΠΏΡΠ΅Π΄Π΅Π»Π΅Π½ΠΈΠ΅ ΡΠ΅Π½ΠΎΡΠΈΠΏΠΎΠ² ΡΡΠ°ΠΌΠΌΠΎΠ² Π΄ΡΠΎΠΆΠΆΠ΅ΠΉ S. cerevisiae
- 3. 2. 8. ΠΠ·ΠΌΠ΅ΡΠ΅Π½ΠΈΠ΅ ΠΏΠΎΠ³Π»ΠΎΡΠ΅Π½ΠΈΡ ΠΊΠΈΡΠ»ΠΎΡΠΎΠ΄Π° ΠΊΠ»Π΅ΡΠΊΠ°ΠΌΠΈ
- 3. 2. 9. ΠΡΠΊΠ»ΡΡΠ΅Π½ΠΈΠ΅ ΠΏΠ»Π°Π·ΠΌΠΈΠ΄Ρ, ΡΠΎΠ΄Π΅ΡΠΆΠ°ΡΠ΅ΠΉ ΠΌΠ°ΡΠΊΠ΅Ρ URA3, ΠΈΠ· Π΄ΡΠΎΠΆΠΆΠ΅Π²ΡΡ ΠΊΠ»Π΅ΡΠΎΠΊ
- 3. 2. 10. ΠΡΠ΄Π΅Π»Π΅Π½ΠΈΠ΅ ΠΠΠ ΠΈΠ· ΠΊΠ»Π΅ΡΠΎΠΊ Π΄ΡΠΎΠΆΠΆΠ΅ΠΉ
- 3. 2. 11. ΠΡΠ΄Π΅Π»Π΅Π½ΠΈΠ΅ ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠΉ ΠΈΠ· ΠΊΠ»Π΅ΡΠΎΠΊ Π΄ΡΠΎΠΆΠΆΠ΅ΠΉ
- 3. 2. 12. ΠΡΠ΄Π΅Π»Π΅Π½ΠΈΠ΅ Π΄ΡΠΎΠΆΠΆΠ΅Π²ΡΡ ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΠ½ΡΡ Π ΠΠ
- 3. 2. 13. ΠΡΠ΄Π΅Π»Π΅Π½ΠΈΠ΅ ΡΡΠΌΠΌΠ°ΡΠ½ΡΡ Π΄ΡΠΎΠΆΠΆΠ΅Π²ΡΡ ΡΠ ΠΠ Π² Π°ΠΌΠΈΠ½ΠΎΠ°ΡΠΈΠ»ΠΈΡΠΎΠ²Π°Π½Π½ΠΎΠΌ ΡΠΎΡΡΠΎΡΠ½ΠΈΠΈ
- 3. 2. 14. ΠΠΎΠ·Π΅ΡΠ½-Π±Π»ΠΎΡ Π³ΠΈΠ±ΡΠΈΠ΄ΠΈΠ·Π°ΡΠΈΡ
- 3. 2. 15. In vitro Π’7-ΡΡΠ°Π½ΡΠΊΡΠΈΠΏΡΠΈΡ
- 3. 2. 16. Π Π°Π΄ΠΈΠΎΠ°ΠΊΡΠΈΠ²Π½ΠΎΠ΅ ΠΌΠ΅ΡΠ΅Π½ΠΈΠ΅ ΡΠ ΠΠ
- 3. 2. 17. ΠΠΊΡΠΏΡΠ΅ΡΡΠΈΡ ΡΠ΅ΠΊΠΎΠΌΠ±ΠΈΠ½Π°Π½ΡΠ½ΡΡ Π±Π΅Π»ΠΊΠΎΠ² Π² ΠΊΠ»Π΅ΡΠΊΠ°Ρ E. col
- 3. 2. 18. ΠΡΠΈΡΡΠΊΠ° ΡΠ΅ΠΊΠΎΠΌΠ±ΠΈΠ½Π°Π½ΡΠ½ΡΡ Π±Π΅Π»ΠΊΠΎΠ² ΠΌΠ΅ΡΠΎΠ΄ΠΎΠΌ Π°ΡΡΠΈΠ½Π½ΠΎΠΉ Ρ ΡΠΎΠΌΠ°ΡΠΎΠ³ΡΠ°ΡΠΈΠΈ Π½Π° ΠΊΠΎΠ»ΠΎΠ½ΠΊΠ°Ρ Ρ Π½ΠΈΠΊΠ΅Π»Ρ-Π°Π³Π°ΡΠΎΠ·ΠΎΠΉ
- 3. 2. 19. ΠΠ΅ΡΡΠ΅ΡΠ½-Π±Π»ΠΎΡ Π³ΠΈΠ±ΡΠΈΠ΄ΠΈΠ·Π°ΡΠΈΡ
- 3. 2. 20. ΠΡΠ΄Π΅Π»Π΅Π½ΠΈΠ΅ Π±Π΅Π»ΠΊΠΎΠ²ΠΎΠ³ΠΎ ΠΏΡΠ΅ΠΏΠ°ΡΠ°ΡΠ°, ΡΠΏΠΎΡΠΎΠ±Π½ΠΎΠ³ΠΎ Π½Π°ΠΏΡΠ°Π²Π»ΡΡΡ ΠΈΠΌΠΏΠΎΡΡ ΡΠ ΠΠ Π² ΠΈΠ·ΠΎΠ»ΠΈΡΠΎΠ²Π°Π½Π½ΡΠ΅ ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠΈ Π΄ΡΠΎΠΆΠΆΠ΅ΠΉ
- 3. 2. 21. Π’ΡΠ°Π½ΡΠΏΠΎΡΡ ΡΠ°Π΄ΠΈΠΎΠ°ΠΊΡΠΈΠ²Π½ΠΎ ΠΌΠ΅ΡΠ΅Π½Π½ΡΡ ΡΠ ΠΠ Π² ΠΈΠ·ΠΎΠ»ΠΈΡΠΎΠ²Π°Π½Π½ΡΠ΅ ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠΈ Π΄ΡΠΎΠΆΠΆΠ΅ΠΉ
- 3. 2. 22. ΠΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΠ½Π°Ρ ΡΡΠ°Π½ΡΠ»ΡΡΠΈΡ in vivo
- 3. 2. 23. ΠΠ΅ΡΠΎΠ΄ Π·Π°Π΄Π΅ΡΠΆΠΊΠΈ Π² Π³Π΅Π»Π΅
- 3. 2. 24. Π€ΡΡΠΏΡΠΈΠ½ΡΠΈΠ½Π³
- 3. 1. ΠΠ°ΡΠ΅ΡΠΈΠ°Π»Ρ
- 4. 1. ΠΠ±ΡΠ°Π·ΠΎΠ²Π°Π½ΠΈΠ΅ Π ΠΠ-Π±Π΅Π»ΠΊΠΎΠ²ΠΎΠ³ΠΎ ΠΊΠΎΠΌΠΏΠ»Π΅ΠΊΡΠ°, ΠΏΡΠΈΠ²ΠΎΠ΄ΡΡΠ΅Π³ΠΎ ΠΊ ΠΈΠΌΠΏΠΎΡΡΡ ΡΠ ΠΠ Π² ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠΈ Π΄ΡΠΎΠΆΠΆΠ΅ΠΉ
- 4. 2. ΠΠ·ΡΡΠ΅Π½ΠΈΠ΅ ΡΡΡΡΠΊΡΡΡΡ Π ΠΠ-Π±Π΅Π»ΠΊΠΎΠ²ΠΎΠ³ΠΎ ΠΊΠΎΠΌΠΏΠ»Π΅ΠΊΡΠ°, ΠΏΡΠΈΠ²ΠΎΠ΄ΡΡΠ΅Π³ΠΎ ΠΊ ΠΈΠΌΠΏΠΎΡΡΡ ΡΠ ΠΠ Π² ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠΈ, ΠΌΠ΅ΡΠΎΠ΄ΠΎΠΌ ΡΡΡΠΏΡΠΈΠ½ΡΠΈΠ½Π³Π°
- 4. 3. ΠΠΎΠΈΡΠΊ ΡΡΠ°ΡΡΠΊΠΎΠ² ΡΠ³Π΅ΠΠ·ΠΊ1Ρ, Π½Π΅ΠΎΠ±Ρ ΠΎΠ΄ΠΈΠΌΡΡ Π΄Π»Ρ Π½Π°ΠΏΡΠ°Π²Π»Π΅Π½ΠΈΡ ΠΈΠΌΠΏΠΎΡΡΠ° ΡΠ ΠΠ Π² ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠΈ
- 4. 4. Π‘ΠΎΠ·Π΄Π°Π½ΠΈΠ΅ ΡΡΠ°ΠΌΠΌΠ° Π΄ΡΠΎΠΆΠΆΠ΅ΠΉ, Π² ΠΊΠΎΡΠΎΡΠΎΠΌ ΡΠ Π1 Π½Π΅ ΠΈΠΌΠΏΠΎΡΡΠΈΡΠΎΠ²Π°Π»Π°ΡΡ Π±Ρ Π² ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠΈ, ΠΈ ΠΈΠ·ΡΡΠ΅Π½ΠΈΠ΅ ΡΠ΅Π½ΠΎΡΠΈΠΏΠΈΡΠ΅ΡΠΊΠΎΠ³ΠΎ ΡΡΡΠ΅ΠΊΡΠ° ΡΠ°ΠΊΠΎΠ³ΠΎ Π΄Π΅ΡΠ΅ΠΊΡΠ°
ΠΠΌΠΏΠΎΡΡ ΡΠ ΠΠ Π² ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠΈ Π΄ΡΠΎΠΆΠΆΠ΅ΠΉ: ΡΠΎΠ»Ρ ΠΏΡΠ΅Π΄ΡΠ΅ΡΡΠ²Π΅Π½Π½ΠΈΠΊΠ° ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΠ½ΠΎΠΉ Π»ΠΈΠ·ΠΈΠ»-ΡΠ ΠΠ-ΡΠΈΠ½ΡΠ΅ΡΠ°Π·Ρ ΠΈ ΡΡΠ½ΠΊΡΠΈΡ ΠΈΠΌΠΏΠΎΡΡΠΈΡΡΠ΅ΠΌΠΎΠΉ ΡΠ ΠΠ Π² ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΠ½ΠΎΠΌ ΠΌΠ°ΡΡΠΈΠΊΡΠ΅ (ΡΠ΅ΡΠ΅ΡΠ°Ρ, ΠΊΡΡΡΠΎΠ²Π°Ρ, Π΄ΠΈΠΏΠ»ΠΎΠΌ, ΠΊΠΎΠ½ΡΡΠΎΠ»ΡΠ½Π°Ρ)
ΠΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠΈ — ΡΡΠΎ ΠΌΠ½ΠΎΠ³ΠΎΡΡΠ½ΠΊΡΠΈΠΎΠ½Π°Π»ΡΠ½ΡΠ΅ ΠΎΡΠ³Π°Π½Π΅Π»Π»Ρ ΡΡΠΊΠ°ΡΠΈΠΎΡΠΈΡΠ΅ΡΠΊΠΈΡ ΠΊΠ»Π΅ΡΠΎΠΊ, ΠΎΠ±Π΅ΡΠΏΠ΅ΡΠΈΠ²Π°ΡΡΠΈΠ΅ ΠΊΠ»Π΅ΡΠΊΠΈ ΡΠ½Π΅ΡΠ³ΠΈΠ΅ΠΉ Π·Π° ΡΡΠ΅Ρ ΠΎΠΊΠΈΡΠ»ΠΈΡΠ΅Π»ΡΠ½ΠΎΠ³ΠΎ ΡΠΎΡΡΠΎΡΠΈΠ»ΠΈΡΠΎΠ²Π°Π½ΠΈΡ, ΡΡΠ°ΡΡΠ²ΡΡΡΠΈΠ΅ Π² ΠΎΠ±ΡΠ°Π·ΠΎΠ²Π°Π½ΠΈΠΈ ΠΆΠ΅Π»Π΅Π·ΠΎΡΠ΅ΡΠ½ΡΡ ΠΊΠ»Π°ΡΡΠ΅ΡΠΎΠ², ΠΎΠΊΠΈΡΠ»Π΅Π½ΠΈΠΈ ΠΆΠΈΡΠ½ΡΡ ΠΊΠΈΡΠ»ΠΎΡ, ΠΎΠ±ΡΠ°Π·ΠΎΠ²Π°Π½ΠΈΠΈ Π½Π΅ΠΊΠΎΡΠΎΡΡΡ Π°ΠΌΠΈΠ½ΠΎΠΊΠΈΡΠ»ΠΎΡ, Π° ΡΠ°ΠΊΠΆΠ΅ ΠΈΠ³ΡΠ°ΡΡΠΈΠ΅ Π²Π°ΠΆΠ½ΡΡ ΡΠΎΠ»Ρ Π² Π°ΠΏΠΎΠΏΡΠΎΠ·Π΅. ΠΠ»Ρ ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠΉ Ρ Π°ΡΠ°ΠΊΡΠ΅ΡΠ½ΠΎ Π½Π°Π»ΠΈΡΠΈΠ΅ ΡΠΎΠ±ΡΡΠ²Π΅Π½Π½ΠΎΠΉ ΠΠΠ ΠΈ ΡΠΈΡΡΠ΅ΠΌΡ Π±ΠΈΠΎΡΠΈΠ½ΡΠ΅Π·Π° Π±Π΅Π»ΠΊΠ°, ΠΎΠ΄Π½Π°ΠΊΠΎ Π±ΠΎΠ»ΡΡΠΎΠ΅ ΡΠΈΡΠ»ΠΎ ΠΌΠ°ΠΊΡΠΎΠΌΠΎΠ»Π΅ΠΊΡΠ» ΠΈΠΌΠΏΠΎΡΡΠΈΡΡΠ΅ΡΡΡ Π² ΠΎΡΠ³Π°Π½Π΅Π»Π»Ρ ΠΈΠ· ΡΠΈΡΠΎΠΏΠ»Π°Π·ΠΌΡ. ΠΡΠ»ΠΎ ΠΏΠΎΠΊΠ°Π·Π°Π½ΠΎ, ΡΡΠΎ Π² ΡΡΠ΄Π΅ ΡΠ»ΡΡΠ°Π΅Π² Π² ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠΈ ΡΡΠ°Π½ΡΠΏΠΎΡΡΠΈΡΡΡΡΡΡ Π½Π΅ ΡΠΎΠ»ΡΠΊΠΎ Π±Π΅Π»ΠΊΠΈ, Π½ΠΎ ΡΠ°ΠΊΠΆΠ΅ ΠΈ ΡΠ ΠΠ. Π Π½Π°ΡΡΠΎΡΡΠ΅ΠΌΡ Π²ΡΠ΅ΠΌΠ΅Π½ΠΈ ΠΈΠΌΠΏΠΎΡΡ ΡΠ ΠΠ Π² ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠΈ Π±ΡΠ» ΠΎΠ±Π½Π°ΡΡΠΆΠ΅Π½ Ρ ΠΏΡΠΎΡΡΠ΅ΠΉΡΠΈΡ , ΡΡΠ΄Π° ΡΠ°ΡΡΠ΅Π½ΠΈΠΉ, ΡΡΠΌΡΠ°ΡΡΡ ΠΌΠ»Π΅ΠΊΠΎΠΏΠΈΡΠ°ΡΡΠΈΡ , Π° ΡΠ°ΠΊΠΆΠ΅ Ρ ΠΏΠΎΡΠΊΡΡΡΠΈΡ ΡΡ Π΄ΡΠΎΠΆΠΆΠ΅ΠΉ Saccharomyces cerevisiae. ΠΠ΅Ρ Π°Π½ΠΈΠ·ΠΌ ΠΏΠ΅ΡΠ΅Π½ΠΎΡΠ° ΡΠ ΠΠ Π² ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠΈ ΠΈ Π΅Π³ΠΎ ΡΠΏΠ΅ΡΠΈΡΠΈΡΠ½ΠΎΡΡΡ Π·Π½Π°ΡΠΈΡΠ΅Π»ΡΠ½ΠΎ ΠΎΡΠ»ΠΈΡΠ°Π΅ΡΡΡ ΠΎΡ Π²ΠΈΠ΄Π° ΠΊ Π²ΠΈΠ΄Ρ.
Π ΡΠ»ΡΡΠ°Π΅ Π΄ΡΠΎΠΆΠΆΠ΅ΠΉ Π±ΡΠ»ΠΎ ΠΏΠΎΠΊΠ°Π·Π°Π½ΠΎ, ΡΡΠΎ ΠΈΠ· ΡΠΈΡΠΎΠΏΠ»Π°Π·ΠΌΡ Π² ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠΈ ΠΈΠΌΠΏΠΎΡΡΠΈΡΡΠ΅ΡΡΡ Π΄Π²Π° Π²ΠΈΠ΄Π° ΡΠ ΠΠ — ΡΠ ΠΠΠΠΈΠ·Π‘ΠΈΠΈ (Martin et al. 1979) ΠΈ ΡΠ ΠΠΠΠ»Π½ (Rinehart et al. 2005). ΠΠ΅ΡΠ°Π»ΡΠ½ΡΠ΅ ΠΌΠ΅Ρ Π°Π½ΠΈΠ·ΠΌΡ ΠΈΠΌΠΏΠΎΡΡΠ° ΡΠ ΠΠ, Π° ΡΠ°ΠΊΠΆΠ΅ ΡΡΠ½ΠΊΡΠΈΠΈ ΠΈΠΌΠΏΠΎΡΡΠΈΡΡΠ΅ΠΌΠΎΠΉ ΡΠ ΠΠ Π² ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΡΠΌ ΠΌΠ°ΡΡΠΈΠΊΡΠ΅, ΠΎΡΡΠ°ΡΡΡΡ Π½Π΅ΡΡΠ½ΡΠΌΠΈ. ΠΠΌΠΏΠΎΡΡ ΡΠ ΠΠ ΡΠ²Π»ΡΠ΅ΡΡΡ Π²ΡΡΠΎΠΊΠΎΡΠΏΠ΅ΡΠΈΡΠΈΡΠ½ΡΠΌ ΠΏΡΠΎΡΠ΅ΡΡΠΎΠΌ, ΡΠ°ΠΊ ΠΊΠ°ΠΊ ΠΎΠ΄Π½Π° ΠΈΠ· Π΄Π²ΡΡ ΠΈΠ·ΠΎΠ°ΠΊΡΠ΅ΠΏΡΠΎΡΠ½ΡΡ Π»ΠΈΠ·ΠΈΠ½ΠΎΠ²ΡΡ ΡΠ ΠΠ (ΡΠ ΠΠΠΠΈΠ·ΡΠΈΠΈ, Π΄Π°Π»Π΅Π΅ ΡΠ Π1) Π½Π°ΠΏΡΠ°Π²Π»ΡΠ΅ΡΡΡ Π² ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠΈ Π΄ΡΠΎΠΆΠΆΠ΅ΠΉ, Π² ΡΠΎ Π²ΡΠ΅ΠΌΡ ΠΊΠ°ΠΊ Π²ΡΠΎΡΠ°Ρ (ΡΠ ΠΠΠΠΈΠ·ΠΈΠΈΠΈ> ΡΠ Π2) ΠΏΡΠΈΡΡΡΡΡΠ²ΡΠ΅Ρ ΡΠΎΠ»ΡΠΊΠΎ Π² ΡΠΈΡΠΎΠΏΠ»Π°Π·ΠΌΠ΅. Π’Π°ΠΊΠΎΠ΅ ΡΠ°Π·Π»ΠΈΡΠΈΠ΅ Π² Π»ΠΎΠΊΠ°Π»ΠΈΠ·Π°ΡΠΈΠΈ Π΄ΠΎΠ²ΠΎΠ»ΡΠ½ΠΎ Π±Π»ΠΈΠ·ΠΊΠΈΡ ΡΠ ΠΠ ΠΌΠΎΠΆΠ΅Ρ Π±ΡΡΡ ΠΎΠ±ΡΡΡΠ½Π΅Π½ΠΎ ΠΎΡΠ»ΠΈΡΠΈΡΠΌΠΈ Π² ΡΡΡΡΠΊΡΡΡΠ΅ ΡΡΠΈΡ ΡΠ ΠΠ (Entelis et al. 1998). ΠΠΎΠΌΠΈΠΌΠΎ ΡΡΠΎΠ³ΠΎ, Π² Π½Π°ΡΠ΅ΠΉ Π»Π°Π±ΠΎΡΠ°ΡΠΎΡΠΈΠΈ Π±ΡΠ»ΠΎ ΠΏΠΎΠΊΠ°Π·Π°Π½ΠΎ ΡΡΠ°ΡΡΠΈΠ΅ ΡΡΠ΅Ρ ΡΠ°ΡΡΠ²ΠΎΡΠΈΠΌΡΡ ΡΠΈΡΠΎΠΏΠ»Π°Π·ΠΌΠ°ΡΠΈΡΠ΅ΡΠΊΠΈΡ Π±Π΅Π»ΠΊΠΎΠ² Π² ΠΈΠΌΠΏΠΎΡΡΠ΅ ΡΠ Π1 Π² ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠΈ. ΠΡΠ΅ΠΆΠ΄Π΅ Π²ΡΠ΅Π³ΠΎ, ΡΠ Π1 Π΄ΠΎΠ»ΠΆΠ½Π° Π±ΡΡΡ Π°ΠΌΠΈΠ½ΠΎΠ°ΡΠΈΠ»ΠΈΡΠΎΠ²Π°Π½Π° Ρ ΠΏΠΎΠΌΠΎΡΡΡ ΡΠΈΡΠΎΠΏΠ»Π°Π·ΠΌΠ°ΡΠΈΡΠ΅ΡΠΊΠΎΠΉ Π»ΠΈΠ·ΠΈΠ»-ΡΠ ΠΠ-ΡΠΈΠ½ΡΠ΅ΡΠ°Π·Ρ, Π·Π°ΡΠ΅ΠΌ Π½Π΅ΠΎΠ±Ρ ΠΎΠ΄ΠΈΠΌΠΎ Π΅Π΅ Π²Π·Π°ΠΈΠΌΠΎΠ΄Π΅ΠΉΡΡΠ²ΠΈΠ΅ Ρ ΠΏΡΠ΅Π΄ΡΠ΅ΡΡΠ²Π΅Π½Π½ΠΈΠΊΠΎΠΌ ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΡΠΉ Π»ΠΈΠ·ΠΈΠ»-ΡΠ ΠΠ-ΡΠΈΠ½ΡΠ΅ΡΠ°Π·Ρ (preMsklp) (Tarassov et al. 1995b). ΠΡΠ΅Π΄ΠΏΠΎΠ»Π°Π³Π°Π΅ΡΡΡ, ΡΡΠΎ ΡΠ Π1 ΠΈΠΌΠΏΠΎΡΡΠΈΡΡΠ΅ΡΡΡ Π² ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠΈ, Π±ΡΠ΄ΡΡΠΈ ΡΠ²ΡΠ·Π°Π½Π½ΠΎΠΉ Ρ ΡΡΠΈΠΌ Π±Π΅Π»ΠΊΠΎΠΌ. ΠΠ΄Π½Π°ΠΊΠΎ ΠΏΡΠΈΡΡΡΡΡΠ²ΠΈΡ Π°ΠΌΠΈΠ½ΠΎΠ°ΡΠΈΠ»ΠΈΡΠΎΠ²Π°Π½Π½ΠΎΠΉ ΡΠ Π1 ΠΈ preMsklp Π½Π΅ Π΄ΠΎΡΡΠ°ΡΠΎΡΠ½ΠΎ Π΄Π»Ρ ΠΈΠΌΠΏΠΎΡΡΠ° ΡΠ Π1 Π² ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠΈ, ΡΡΠΎ Π³ΠΎΠ²ΠΎΡΠΈΡ ΠΎ Π½Π΅ΠΎΠ±Ρ ΠΎΠ΄ΠΈΠΌΠΎΡΡΠΈ Π΄ΠΎΠΏΠΎΠ»Π½ΠΈΡΠ΅Π»ΡΠ½ΡΡ Π±Π΅Π»ΠΊΠΎΠ²ΡΡ ΡΠ°ΠΊΡΠΎΡΠΎΠ², Π½Π°ΠΏΡΠ°Π²Π»ΡΡΡΠΈΡ Π΅Π΅ ΠΈΠΌΠΏΠΎΡΡ, ΡΠ°ΠΊ Π½Π°Π·ΡΠ²Π°Π΅ΠΌΡΡ ΡΠ°ΠΊΡΠΎΡΠΎΠ² ΠΈΠΌΠΏΠΎΡΡΠ°. ΠΠ΄ΠΈΠ½ ΠΈΠ· ΡΠ°ΠΊΠΈΡ ΡΠ°ΠΊΡΠΎΡΠΎΠ² Π±ΡΠ» Π½Π΅Π΄Π°Π²Π½ΠΎ ΠΎΡ Π°ΡΠ°ΠΊΡΠ΅ΡΠΈΠ·ΠΎΠ²Π°Π½ ΠΊΠ°ΠΊ Π³Π»ΠΈΠΊΠΎΠ»ΠΈΡΠΈΡΠ΅ΡΠΊΠΈΠΉ ΡΠ΅ΡΠΌΠ΅Π½Ρ Π΅Π½ΠΎΠ»Π°Π·Π° (Entelis et al. 2006). Π ΠΎΠ»Ρ ΡΡΠΎΠ³ΠΎ Π±Π΅Π»ΠΊΠ° Π² ΠΏΡΠΎΡΠ΅ΡΡΠ΅ ΠΈΠΌΠΏΠΎΡΡΠ° ΡΠ ΠΠ Π² ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠΈ ΠΎΡΡΠ°Π΅ΡΡΡ Π½Π΅ΡΡΠ½ΠΎΠΉ.
ΠΠΎΠΏΡΠΎΡ ΠΎ ΡΡΠ½ΠΊΡΠΈΠΈ tPJII Π² ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΠ½ΠΎΠΌ ΠΌΠ°ΡΡΠΈΠΊΡΠ΅ ΠΊ Π½Π°ΡΡΠΎΡΡΠ΅ΠΌΡ ΠΌΠΎΠΌΠ΅Π½ΡΡ ΠΎΡΡΠ°Π΅ΡΡΡ ΠΎΡΠΊΡΡΡΡΠΌ. Π‘ΡΡΠ΅ΡΡΠ²ΡΠ΅Ρ ΠΏΡΠ΅Π΄ΠΏΠΎΠ»ΠΎΠΆΠ΅Π½ΠΈΠ΅, ΡΡΠΎ ΡΠ Π1 ΠΏΡΠΈΠ½ΠΈΠΌΠ°Π΅Ρ ΡΡΠ°ΡΡΠΈΠ΅ Π² ΠΌΠΈΡΠΎΡ ΠΎΠΈΠ΄ΡΠΈΠ°Π»ΡΠ½ΠΎΠΉ ΡΡΠ°Π½ΡΠ»ΡΡΠΈΠΈ. ΠΠ΄Π½Π°ΠΊΠΎ, ΡΡΠ° Π³ΠΈΠΏΠΎΡΠ΅Π·Π° ΠΏΠΎΠ΄ΡΠ²Π΅ΡΠΆΠ΄Π΅Π½Π° ΡΠΎΠ»ΡΠΊΠΎ ΠΊΠΎΡΠ²Π΅Π½Π½ΡΠΌΠΈ ΡΠΊΡΠΏΠ΅ΡΠΈΠΌΠ΅Π½ΡΠ°ΠΌΠΈ (Kolesnikova et al. 2000). ΠΠΎΠ»Π΅Π΅ ΡΠΎΠ³ΠΎ, ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΠ½Π°Ρ Π»ΠΈΠ·ΠΈΠ½ΠΎΠ²Π°Ρ ΡΠ ΠΠ Π΄ΡΠΎΠΆΠΆΠ΅ΠΉ (Ρ Π°Π½ΡΠΈΠΊΠΎΠ΄ΠΎΠ½ΠΎΠΌ cmnmsUUU, Π΄Π°Π»Π΅Π΅ ΡΠ ΠΠ) ΡΠΏΠΎΡΠΎΠ±Π½Π° ΡΠ·Π½Π°Π²Π°ΡΡ ΠΎΠ±Π° Π»ΠΈΠ·ΠΈΠ½ΠΎΠ²ΡΡ ΠΊΠΎΠ΄ΠΎΠ½Π° (AAA ΠΈ AAG) Π² ΠΏΡΠΎΡΠ΅ΡΡΠ΅ ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΡΠΉ ΡΡΠ°Π½ΡΠ»ΡΡΠΈΠΈ.
Π¦Π΅Π»ΡΡ Π΄Π°Π½Π½ΠΎΠΉ ΡΠ°Π±ΠΎΡΡ ΡΠ²Π»ΡΠ»ΠΎΡΡ ΠΈΠ·ΡΡΠ΅Π½ΠΈΠ΅ ΡΠΎΡΠΌΠΈΡΠΎΠ²Π°Π½ΠΈΡ ΠΊΠΎΠΌΠΏΠ»Π΅ΠΊΡΠ° ΠΌΠ΅ΠΆΠ΄Ρ ΡΠ Π1 ΠΈ preMsklp, ΠΈ Ρ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠ΅ΠΌ ΠΏΠΎΠ»ΡΡΠ΅Π½Π½ΡΡ Π΄Π°Π½Π½ΡΡ , ΡΠΎΠ·Π΄Π°Π½ΠΈΠ΅ ΡΡΠ°ΠΌΠΌΠ° Π΄ΡΠΎΠΆΠΆΠ΅ΠΉ, Π² ΠΊΠΎΡΠΎΡΠΎΠΌ ΡΠ Π1 ΠΏΡΠΈΡΡΡΡΡΠ²ΠΎΠ²Π°Π»Π° Π±Ρ ΠΈΡΠΊΠ»ΡΡΠΈΡΠ΅Π»ΡΠ½ΠΎ Π² ΡΠΈΡΠΎΠΏΠ»Π°Π·ΠΌΠ΅, ΠΈ ΠΈΠ·ΡΡΠ΅Π½ΠΈΠ΅ ΡΠ΅Π½ΠΎΡΠΈΠΏΠΈΡΠ΅ΡΠΊΠΎΠ³ΠΎ ΡΡΡΠ΅ΠΊΡΠ° ΠΎΡΡΡΡΡΡΠ²ΠΈΡ ΡΠ Π1 Π² ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΡΡ .
ΠΠ»Ρ Π΄ΠΎΡΡΠΈΠΆΠ΅Π½ΠΈΡ ΠΏΠΎΡΡΠ°Π²Π»Π΅Π½Π½ΡΡ ΡΠ΅Π»Π΅ΠΉ Π±ΡΠ»ΠΈ ΡΡΠΎΡΠΌΡΠ»ΠΈΡΠΎΠ²Π°Π½Ρ ΡΠ»Π΅Π΄ΡΡΡΠΈΠ΅ ΡΠΊΡΠΏΠ΅ΡΠΈΠΌΠ΅Π½ΡΠ°Π»ΡΠ½ΡΠ΅ Π·Π°Π΄Π°ΡΠΈ:
— ΠΎΡΠ΅Π½ΠΈΡΡ ΠΊΠΎΠ½ΡΡΠ°Π½ΡΡ Π΄ΠΈΡΡΠΎΡΠΈΠ°ΡΠΈΠΈ ΠΊΠΎΠΌΠΏΠ»Π΅ΠΊΡΠ° ΡΠ Π1-preMsklp Π² ΠΎΡΡΡΡΡΡΠ²ΠΈΠΈ ΠΈ ΠΏΡΠΈΡΡΡΡΡΠ²ΠΈΠΈ Π΅Π½ΠΎΠ»Π°Π·Ρ 2;
— ΠΎΠΏΡΠ΅Π΄Π΅Π»ΠΈΡΡ, ΠΊΠ°ΠΊΠΈΠ΅ Π½ΡΠΊΠ»Π΅ΠΎΡΠΈΠ΄Ρ ΡΠ Π1 ΡΡΠ°Π½ΠΎΠ²ΡΡΡΡ Π·Π°ΡΠΈΡΠ΅Π½Π½ΡΠΌΠΈ ΠΎΡ Π΄Π΅ΠΉΡΡΠ²ΠΈΡ Π½ΡΠΊΠ»Π΅Π°Π· Π² ΡΠ΅Π·ΡΠ»ΡΡΠ°ΡΠ΅ Π²Π·Π°ΠΈΠΌΠΎΠ΄Π΅ΠΉΡΡΠ²ΠΈΡ Ρ preMsklp;
— ΠΎΡΠ΅ΠΏΠΈΡΡ ΡΡΡΠ΅ΠΊΡΠΈΠ²Π½ΠΎΡΡΡ ΠΈΠΌΠΏΠΎΡΡΠ° ΡΠ Π1, Π½Π°ΠΏΡΠ°Π²Π»ΡΠ΅ΠΌΠΎΠ³ΠΎ ΡΠ°Π·Π»ΠΈΡΠ½ΡΠΌΠΈ ΠΌΡΡΠ°Π½ΡΠ½ΡΠΌΠΈ Π²Π΅ΡΡΠΈΡΠΌΠΈ preMsklp in vitro ΠΈ in vivo;
— Π² ΡΠ»ΡΡΠ°Π΅ Π½Π΅Π²ΠΎΠ·ΠΌΠΎΠΆΠ½ΠΎΡΡΠΈ Π½Π°ΠΏΡΠ°Π²Π»Π΅Π½ΠΈΡ ΠΈΠΌΠΏΠΎΡΡΠ° ΡΠ Π1 ΡΠΎΠΉ ΠΈΠ»ΠΈ ΠΈΠ½ΠΎΠΉ ΠΌΡΡΠ°Π½ΡΠ½ΠΎΠΉ Π²Π΅ΡΡΠΈΠ΅ΠΉ preMsklp, ΠΎΡΠ΅Π½ΠΈΡΡ ΡΡΡΠ΅ΠΊΡΠΈΠ²Π½ΠΎΡΡΡ ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΠ½ΠΎΠ³ΠΎ Π΄ΡΡ Π°Π½ΠΈΡ ΠΈ ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΡΠΉ ΡΡΠ°Π½ΡΠ»ΡΡΠΈΠΈ Π² ΠΊΠ»Π΅ΡΠΊΠ°Ρ ΡΡΠ°ΠΌΠΌΠ° Π΄ΡΠΎΠΆΠΆΠ΅ΠΉ Ρ ΡΠΎΠΎΡΠ²Π΅ΡΡΡΠ²ΡΡΡΠ΅ΠΉ ΠΌΡΡΠ°Π½ΡΠ½ΠΎΠΉ Π²Π΅ΡΡΠΈΠ΅ΠΉ preMsklp;
— ΠΎΡΠ΅Π½ΠΈΡΡ ΡΡΡΠ΅ΠΊΡΠΈΠ²Π½ΠΎΡΡΡ ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΠ½ΠΎΠ³ΠΎ Π΄ΡΡ Π°Π½ΠΈΡ ΠΈ ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΡΠΉ ΡΡΠ°Π½ΡΠ»ΡΡΠΈΠΈ Π² ΠΊΠ»Π΅ΡΠΊΠ°Ρ ΡΡΠ°ΠΌΠΌΠ° Π΄ΡΠΎΠΆΠΆΠ΅ΠΉ, Π² ΠΊΠΎΡΠΎΡΡΡ Π³Π΅ΠΈ preMsklp Π·Π°ΠΌΠ΅Π½Π΅Π½ Π½Π° ΠΎΡΡΠΎΠ»ΠΎΠ³ΠΈΡΠ½ΡΠΉ Π³Π΅Π½ Π³ΡΠΈΠ±ΠΎΠ² Ashbya gossypii.
5. ΠΠ«ΠΠΠΠ«.
1. Π’ΠΎΠ»ΡΠΊΠΎ ΡΠ΅ ΡΠ ΠΠ, ΠΊΠΎΡΠΎΡΡΠ΅ ΡΠΏΠΎΡΠΎΠ±Π½Ρ ΠΎΠ±ΡΠ°Π·ΠΎΠ²ΡΠ²Π°ΡΡ ΡΡΠ°Π±ΠΈΠ»ΡΠ½ΡΠΉ ΠΊΠΎΠΌΠΏΠ»Π΅ΠΊΡ Ρ ΠΏΡΠ΅Π΄ΡΠ΅ΡΡΠ²Π΅Π½Π½ΠΈΠΊΠΎΠΌ ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΡΠΉ Π»ΠΈΠ·ΠΈΠ»-ΡΠ ΠΠ-ΡΠΈΠ½ΡΠ΅ΡΠ°Π·Ρ (preMsklp), ΠΈΠΌΠΏΠΎΡΡΠΈΡΡΡΡΡΡ Π² ΠΈΠ·ΠΎΠ»ΠΈΡΠΎΠ²Π°Π½Π½ΡΠ΅ ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠΈ Π΄ΡΠΎΠΆΠΆΠ΅ΠΉ. ΠΠΎΠ½ΡΡΠ°Π½ΡΠ° Π΄ΠΈΡΡΠΎΡΠΈΠ°ΡΠΈΠΈ ΡΠ°ΠΊΠΈΡ ΠΊΠΎΠΌΠΏΠ»Π΅ΠΊΡΠΎΠ² ΡΠΎΡΡΠ°Π²Π»ΡΠ΅Ρ 180−250 Π½Π. ΠΡΠΈΡΡΡΡΡΠ²ΠΈΠ΅ Π΅Π½ΠΎΠ»Π°Π·Ρ ΡΠ½ΠΈΠΆΠ°Π΅Ρ ΠΊΠΎΠ½ΡΡΠ°Π½ΡΡ Π΄ΠΈΡΡΠΎΡΠΈΠ°ΡΠΈΠΈ Π΄ΠΎ 40 Π½Π.
2. ΠΠΎΠΌΠΏΠ»Π΅ΠΊΡ ΠΈΠΌΠΏΠΎΡΡΠΈΡΡΠ΅ΠΌΠΎΠΉ Π»ΠΈΠ·ΠΈΠ½ΠΎΠ²ΠΎΠΉ ΡΠ ΠΠ (ΡΠ Π1) Ρ ΡΠΈΡΠΎΠΏΠ»Π°Π·ΠΌΠ°ΡΠΈΡΠ΅ΡΠΊΠΎΠΉ Π»ΠΈΠ·ΠΈΠ»-ΡΠ ΠΠ-ΡΠΈΠ½ΡΠ΅ΡΠ°Π·ΠΎΠΉ ΠΎΡΠ³Π°Π½ΠΈΠ·ΠΎΠ²Π°Π½ ΠΊΠ°ΠΊ ΡΠΈΠΏΠΈΡΠ½ΡΠΉ ΠΊΠΎΠΌΠΏΠ»Π΅ΠΊΡ Π°ΠΌΠΈΠ½ΠΎΠ°ΡΠΈΠ»-ΡΠ ΠΠ-ΡΠΈΠ½ΡΠ΅ΡΠ°Π·Ρ ΠΊΠ»Π°ΡΡΠ° ΠΠ¬ Ρ ΡΠΎΠΎΡΠ²Π΅ΡΡΡΠ²ΡΡΡΠ΅ΠΉ ΡΠ ΠΠ, ΠΎΠ±ΡΠ°Π·ΠΎΠ²Π°Π½ΠΈΠ΅ ΠΊΠΎΡΠΎΡΠΎΠ³ΠΎ Π²Π΅Π΄Π΅Ρ ΠΊ Π°ΠΌΠΈΠ½ΠΎΠ°ΡΠΈΠ»ΠΈΡΠΎΠ²Π°Π½ΠΈΡ. ΠΠΎΠΌΠΏΠ»Π΅ΠΊΡ ΡΠ Π1 Ρ preMsklp ΠΎΡΠ³Π°Π½ΠΈΠ·ΠΎΠ²Π°Π½ ΡΠΎΠ²Π΅ΡΡΠ΅Π½Π½ΠΎ ΠΈΠ½Π°ΡΠ΅: Π² ΡΡΠΎΠΌ ΠΊΠΎΠΌΠΏΠ»Π΅ΠΊΡΠ΅ ΠΏΠΎΠ΄Π°Π²Π»ΡΡΡΠ΅Π΅ Π±ΠΎΠ»ΡΡΠΈΠ½ΡΡΠ²ΠΎ Π½ΡΠΊΠ»Π΅ΠΎΡΠΈΠ΄ΠΎΠ² ΡΠ ΠΠ ΡΠΊΡΠ°Π½ΠΈΡΠΎΠ²Π°Π½Ρ Π±Π΅Π»ΠΊΠΎΠΌ.
3. N-ΠΊΠΎΠ½ΡΠ΅Π²ΠΎΠΉ Π΄ΠΎΠΌΠ΅Π½ preMsklp ΡΠΏΠ΅ΡΠΈΡΠΈΡΠ΅ΡΠΊΠΈ Π½Π°ΠΏΡΠ°Π²Π»ΡΠ΅Ρ ΠΈΠΌΠΏΠΎΡΡ ΡΠ ΠΠ Π² ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠΈ Π΄ΡΠΎΠΆΠΆΠ΅ΠΉ in vitro ΠΈ in vivo ΠΈ Π²Π·Π°ΠΈΠΌΠΎΠ΄Π΅ΠΉΡΡΠ²ΡΠ΅Ρ Ρ ΡΠ Π1 ΡΠ°ΠΊΠΈΠΌ ΠΆΠ΅ ΠΎΠ±ΡΠ°Π·ΠΎΠΌ, ΡΡΠΎ ΠΈ ΠΏΠΎΠ»Π½ΠΎΡΠ°Π·ΠΌΠ΅ΡΠ½ΡΠΉ Π±Π΅Π»ΠΎΠΊ.
Π£ΡΠ°ΡΡΠΎΠΊ preMsklp, ΠΎΠΏΡΠ΅Π΄Π΅Π»ΡΡΡΠΈΠΉ ΡΠΏΠ΅ΡΠΈΡΠΈΡΠ΅ΡΠΊΠΈΠΉ ΠΈΠΌΠΏΠΎΡΡ ΡΠ ΠΠ Π² ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠΈ, ΠΎΠ³ΡΠ°Π½ΠΈΡΠ΅Π½ a-ΡΠΏΠΈΡΠ°Π»ΡΠΌΠΈ Π5 ΠΈ Π6. Π£ΡΠ°ΡΡΠΊΠΈ preMsklp, ΠΎΠ³ΡΠ°Π½ΠΈΡΠ΅Π½Π½ΡΠ΅ a-ΡΠΏΠΈΡΠ°Π»ΡΠΌΠΈ Π5 ΠΈ Π6 ΠΈ a-ΡΠΏΠΈΡΠ°Π»ΡΠΌΠΈ HI ΠΈ ΠΠ, Π½Π΅ΠΎΠ±Ρ ΠΎΠ΄ΠΈΠΌΡ Π΄Π»Ρ Π°ΠΌΠΈΠ½ΠΎΠ°ΡΠΈΠ»ΠΈΡΠΎΠ²Π°Π½ΠΈΡ Π·ΡΠ΅Π»ΡΠΌ Π±Π΅Π»ΠΊΠΎΠΌ ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΠ½ΠΎΠΉ Π»ΠΈΠ·ΠΈΠ½ΠΎΠ²ΠΎΠΉ ΡΠ ΠΠ.
4. ΡΠ Π1 ΡΡΠ°ΡΡΠ²ΡΠ΅Ρ Π² ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΠ½ΠΎΠΉ ΡΡΠ°Π½ΡΠ»ΡΡΠΈΠΈ. ΠΡΠΈ 30 Β°C ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΠ½Π°Ρ Π»ΠΈΠ·ΠΈΠΏΠΎΠ²Π°Ρ ΡΠ ΠΠ (Ρ Π°Π½ΡΠΈΠΊΠΎΠ΄ΠΎΠ½ΠΎΠΌ UUU) ΠΏΠΎΠ»Π½ΠΎΡΡΡΡ Π·Π°ΠΌΠ΅Π½ΡΠ΅Ρ Π² ΡΡΠΎΠΌ ΠΏΡΠΎΡΠ΅ΡΡΠ΅ ΡΠ Π1 (Ρ Π°Π½ΡΠΈΠΊΠΎΠ΄ΠΎΠ½ΠΎΠΌ CUU), ΡΠΎΠ³Π΄Π° ΠΊΠ°ΠΊ ΠΏΡΠΈ 37 Β°C ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΠ½Π°Ρ Π»ΠΈΠ·ΠΈΠ½ΠΎΠ²Π°Ρ ΡΠ ΠΠ Π³ΠΈΠΏΠΎΠΌΠΎΠ΄ΠΈΡΠΈΡΠΈΡΠΎΠ²Π°Π½Π° ΠΏΠΎ U34 ΠΈ Π½Π΅ΡΡΡΠ΅ΠΊΡΠΈΠ²Π½ΠΎ ΡΡΠΈΡΡΠ²Π°Π΅Ρ ΠΊΠΎΠ΄ΠΎΠ½Ρ AAG. ΠΠΎΡΡΠΎΠΌΡ ΠΏΡΠΈ Π΄Π°Π½Π½ΠΎΠΉ ΡΠ΅ΠΌΠΏΠ΅ΡΠ°ΡΡΡΠ΅ ΡΠ Π1 Π½Π΅ΠΎΠ±Ρ ΠΎΠ΄ΠΈΠΌΠ° Π΄Π»Ρ Π½ΠΎΡΠΌΠ°Π»ΡΠ½ΠΎΠ³ΠΎ ΡΡΠ½ΠΊΡΠΈΠΎΠ½ΠΈΡΠΎΠ²Π°Π½ΠΈΡ ΠΌΠΈΡΠΎΡ ΠΎΠ½Π΄ΡΠΈΠΉ.
ΠΠΠΠΠΠΠΠ ΠΠΠ‘Π’Π.
ΠΠ²ΡΠΎΡ ΡΡΠΈΡΠ°Π΅Ρ ΡΠ²ΠΎΠΈΠΌ Π΄ΠΎΠ»Π³ΠΎΠΌ Π²ΡΡΠ°Π·ΠΈΡΡ Π³Π»ΡΠ±ΠΎΡΠ°ΠΉΡΡΡ ΠΏΡΠΈΠ·Π½Π°ΡΠ΅Π»ΡΠ½ΠΎΡΡΡ ΠΈ ΠΎΠ³ΡΠΎΠΌΠ½ΡΡ Π±Π»Π°Π³ΠΎΠ΄Π°ΡΠ½ΠΎΡΡΡ ΡΠ²ΠΎΠΈΠΌ Π½Π°ΡΡΠ½ΡΠΌ ΡΡΠΊΠΎΠ²ΠΎΠ΄ΠΈΡΠ΅Π»ΡΠΌ, ΠΠ³ΠΎΡΡ ΠΠ»Π΅ΠΊΡΠ°Π½Π΄ΡΠΎΠ²ΠΈΡΡ ΠΡΠ°ΡΠ΅Π½ΠΈΠ½Π½ΠΈΠΊΠΎΠ²Ρ ΠΈ ΠΠ²Π°Π½Ρ Π’Π°ΡΠ°ΡΠΎΠ²Ρ. ΠΠ½ΠΈ Π²ΡΠ΅ΠΌΠ΅ΡΠ½ΠΎ ΠΏΠΎΠΌΠΎΠ³Π°Π»ΠΈ ΠΌΠ½Π΅ ΠΏΡΠΈ ΠΏΠΎΡΡΠ°Π½ΠΎΠ²ΠΊΠ΅ ΡΠΊΡΠΏΠ΅ΡΠΈΠΌΠ΅Π½ΡΠΎΠ², ΠΈΠ½ΡΠ΅ΡΠΏΡΠ΅ΡΠ°ΡΠΈΠΈ ΡΠ΅Π·ΡΠ»ΡΡΠ°ΡΠΎΠ² ΠΈ Π½Π°ΠΏΠΈΡΠ°Π½ΠΈΠΈ ΡΡΠ°ΡΠ΅ΠΉ ΠΈ Π΄ΠΈΡΡΠ΅ΡΡΠ°ΡΠΈΠΈ. ΠΠΎΠ»Π΅Π΅ ΡΠΎΠ³ΠΎ, ΠΏΠΎ ΠΏΡΠΎΡΠ΅ΡΡΠ²ΠΈΠΈ ΡΠ΅ΠΌΠΈ Π»Π΅Ρ ΡΠΎΠ²ΠΌΠ΅ΡΡΠ½ΠΎΠΉ ΡΠ°Π±ΠΎΡΡ Ρ ΡΠΌΠ΅Π»ΠΎ ΠΌΠΎΠ³Ρ Π½Π°Π·Π²Π°ΡΡ ΠΈΡ ΡΠ²ΠΎΠΈΠΌΠΈ Π΄ΡΡΠ·ΡΡΠΌΠΈ, Ρ ΠΊΠΎΡΠΎΡΡΠΌΠΈ ΠΌΠ½Π΅ Π²ΡΠ΅Π³Π΄Π° ΠΎΡΠ΅Π½Ρ ΠΈΠ½ΡΠ΅ΡΠ΅ΡΠ½ΠΎ ΠΏΡΠΎΠ²ΠΎΠ΄ΠΈΡΡ Π²ΡΠ΅ΠΌΡ ΠΈ ΠΊΠΎΡΠΎΡΡΠ΅ Π½Π΅ ΡΠ°Π· ΠΏΡΠΈΡ ΠΎΠ΄ΠΈΠ»ΠΈ ΠΌΠ½Π΅ Π½Π° ΠΏΠΎΠΌΠΎΡΡ Π² ΡΡΡΠ΄Π½ΡΡ ΠΌΠΈΠ½ΡΡΡ. ΠΡΠΈ ΠΆΠ΅ ΡΠ»ΠΎΠ²Π° Ρ Ρ ΠΎΡΠ΅Π» Π±Ρ Π°Π΄ΡΠ΅ΡΠΎΠ²Π°ΡΡ ΡΠ²ΠΎΠΈΠΌ ΠΊΠΎΠ»Π»Π΅Π³Π°ΠΌ, ΠΠΈΠ½Π΅ ΠΠ½ΡΠ΅Π»ΠΈΡ, Π ΠΎΠ±ΡΡΡ ΠΠ°ΡΡΡΠ½Ρ ΠΈ ΠΠ»ΡΠ³Π΅ ΠΠΎΠ»Π΅ΡΠ½ΠΈΠΊΠΎΠ²ΠΎΠΉ, Π±Π΅Π· ΠΏΠΎΠΌΠΎΡΠΈ ΠΈ ΠΏΠΎΠ΄Π΄Π΅ΡΠΆΠΊΠΈ ΠΊΠΎΡΠΎΡΡΡ ΡΡΠ° ΡΠ°Π±ΠΎΡΠ° Π½ΠΈΠΊΠΎΠ³Π΄Π° Π½Π΅ Π±ΡΠ»Π° Π±Ρ ΡΠ΄Π΅Π»Π°Π½Π°. Π― ΠΎΡΠ΅Π½Ρ ΠΏΡΠΈΠ·Π½Π°ΡΠ΅Π»Π΅Π½ ΠΡΠΈΠ½Π΅ ΠΡΠ°Π½Π΄ΠΈΠ½ΠΎΠΉ, ΠΠ»ΡΠ³Π΅ ΠΠ°ΡΠΈΡΠ΅Π²ΠΎΠΉ ΠΈ ΠΠ»Π΅ΠΊΡΠ°Π½Π΄ΡΡ Π‘ΠΌΠΈΡΠ½ΠΎΠ²Ρ, ΠΊΠΎΡΠΎΡΡΠ΅, Π² ΡΠΈΡΠ»Π΅ ΠΏΡΠΎΡΠΈΡ , ΡΠΎΠ·Π΄Π°Π»ΠΈ Π² Π½Π°ΡΠ΅ΠΉ Π»Π°Π±ΠΎΡΠ°ΡΠΎΡΠΈΠΈ ΠΈΠ΄Π΅Π°Π»ΡΠ½ΡΠΉ ΠΊΠ»ΠΈΠΌΠ°Ρ Π΄Π»Ρ ΠΏΡΠΈΡΡΠ½ΠΎΠΉ ΠΈ ΡΡΡΠ΅ΠΊΡΠΈΠ²Π½ΠΎΠΉ ΡΠ°Π±ΠΎΡΡ. Π― Π±Π»Π°Π³ΠΎΠ΄Π°ΡΡ ΡΠ°ΠΊΠΆΠ΅ ΡΡΡΠ΄Π΅Π½ΡΠΎΠ², Π΄Π΅Π»Π°Π²ΡΠΈΡ ΠΊΡΡΡΠΎΠ²ΡΠ΅ ΠΈ Π΄ΠΈΠΏΠ»ΠΎΠΌΠ½ΡΠ΅ ΡΠ°Π±ΠΎΡΡ (ΠΈΠ»ΠΈ ΠΈΡ ΡΡΠ°Π½ΡΡΠ·ΡΠΊΠΈΠ΅ Π°Π½Π°Π»ΠΎΠ³ΠΈ) Π² Π½Π°ΡΠ΅ΠΉ Π»Π°Π±ΠΎΡΠ°ΡΠΎΡΠΈΠΈ — ΠΠ»ΠΈΠ½Ρ ΠΠΎΡΠ±ΡΡ, ΠΡΡΠ΅Π»Ρ ΠΡΠΈΡΡΠ΅Ρ, Π‘ΡΠΌΠΈ ΠΠΎΠ³Π°Π½Π° ΠΈ ΠΠ°Π½Π΅ΡΡΡ ΠΡΠ±Π΅Π½ΠΎ. Π§Π°ΡΡΡ ΡΡΠΎΠΉ ΡΠ°Π±ΠΎΡΡ Π±ΡΠ»Π° ΡΠ΄Π΅Π»Π°Π½Π° ΠΏΡΠΈ ΠΈΡ Π½Π΅ΠΏΠΎΡΡΠ΅Π΄ΡΡΠ²Π΅Π½Π½ΠΎΠΌ ΡΡΠ°ΡΡΠΈΠΈ. Π― ΡΠ°ΠΊΠΆΠ΅ Ρ ΠΎΡΠ΅Π» Π±Ρ ΠΏΠΎΠ±Π»Π°Π³ΠΎΠ΄Π°ΡΠΈΡΡ ΡΡΠ΅Π½ΡΡ , ΠΊΠΎΡΠΎΡΡΠ΅ Π»ΡΠ±Π΅Π·Π½ΠΎ Π΄Π΅Π»ΠΈΠ»ΠΈΡΡ ΡΠΎ ΠΌΠ½ΠΎΠΉ Π³Π΅Π½Π½ΠΎ-ΠΈΠ½ΠΆΠ΅Π½Π΅ΡΠ½ΡΠΌΠΈ ΠΊΠΎΠ½ΡΡΡΡΠΊΡΠΈΡΠΌΠΈ, Π°Π½ΡΠΈΡΠ΅Π»Π°ΠΌΠΈ ΠΈ ΠΌΠ΅ΡΠΎΠ΄ΠΈΠΊΠ°ΠΌΠΈ: Π‘ΠΈΠ»ΡΠ²ΠΈ Π€ΡΠΈΠ°Π½, Π£Π±Π΅ΡΠ° ΠΠ΅ΠΊΠΊΠ΅ΡΠ°, ΠΠ»Π΅ΠΊΡΠ°Π½Π΄Π΅ΡΠ° Π¦Π°Π³ΠΎΠ»ΠΎΡΡ, Π₯Π°ΠΉΠΊΠ΅ ΠΠ°Π½Π³ ΠΈ Π ΠΎΠ»Π°Π½Π΄Π° ΠΠΈΠ»Π»Ρ. Π― ΠΎΡΠ΅Π½Ρ ΠΏΡΠΈΠ·Π½Π°ΡΠ΅Π»Π΅Π½ ΠΊΠΎΠ»Π»Π΅ΠΊΡΠΈΠ²Ρ ΠΊΠ°ΡΠ΅Π΄ΡΡ ΠΌΠΎΠ»Π΅ΠΊΡΠ»ΡΡΠ½ΠΎΠΉ Π±ΠΈΠΎΠ»ΠΎΠ³ΠΈΠΈ Π·Π° ΠΏΠΎΠ»ΡΡΠ΅Π½Π½ΠΎΠ΅ ΠΌΠ½ΠΎΠΉ Π²Π΅Π»ΠΈΠΊΠΎΠ»Π΅ΠΏΠ½ΠΎΠ΅ ΠΎΠ±ΡΠ°Π·ΠΎΠ²Π°Π½ΠΈΠ΅. Π’Π°ΠΊΠΆΠ΅ Ρ ΠΎΡΠ΅Π»ΠΎΡΡ Π±Ρ ΠΏΠΎΠ±Π»Π°Π³ΠΎΠ΄Π°ΡΠΈΡΡ ΠΊΠΎΠ»Π»Π΅ΠΊΡΠΈΠ² Π»Π°Π±ΠΎΡΠ°ΡΠΎΡΠΈΠΈ ΠΌΠΎΠ»Π΅ΠΊΡΠ»ΡΡΠ½ΠΎΠΉ Π³Π΅Π½Π΅ΡΠΈΠΊΠΈ, Π³Π΅Π½ΠΎΠΌΠΈΠΊΠΈ ΠΈ ΠΌΠΈΠΊΡΠΎΠ±ΠΈΠΎΠ»ΠΎΠ³ΠΈΠΈ Π£Π½ΠΈΠ²Π΅ΡΡΠΈΡΠ΅ΡΠ° ΠΡΠΈ ΠΠ°ΡΡΠ΅ΡΠ°, Π‘ΡΡΠ°ΡΠ±ΡΡΠ³, Π€ΡΠ°Π½ΡΠΈΡ, ΠΈ Π² ΠΎΡΠΎΠ±Π΅Π½Π½ΠΎΡΡΠΈ — ΠΠ½Π½-ΠΠ°ΡΠΈ ΠΠΊΠ΅Π»Ρ ΠΈ ΠΠ°ΡΠΈ Π Π΅ΠΉΠ±Π΅Π»Ρ, Π·Π° ΠΏΠΎΠΌΠΎΡΡ Π² ΡΠ°Π±ΠΎΡΠ΅ ΠΈ Π΄ΠΎΠ±ΡΠΎΠΆΠ΅Π»Π°ΡΠ΅Π»ΡΠ½ΠΎΠ΅ ΠΎΡΠ½ΠΎΡΠ΅Π½ΠΈΠ΅. Π Π² Π·Π°ΠΊΠ»ΡΡΠ΅Π½ΠΈΠ΅ Ρ Π±Π»Π°Π³ΠΎΠ΄Π°ΡΡ ΡΠ²ΠΎΠΈΡ ΡΠΎΠ΄ΠΈΡΠ΅Π»Π΅ΠΉ ΠΈ ΡΠ²ΠΎΡ ΠΆΠ΅Π½Ρ Π·Π° Π²ΡΠ΅ ΡΠΎ, ΡΡΠΎ ΠΎΠ½ΠΈ Π΄Π»Ρ ΠΌΠ΅Π½Ρ ΡΠ΄Π΅Π»Π°Π»ΠΈ ΠΈ ΠΏΡΠΎΠ΄ΠΎΠ»ΠΆΠ°ΡΡ Π΄Π΅Π»Π°ΡΡ.
Π‘ΠΏΠΈΡΠΎΠΊ Π»ΠΈΡΠ΅ΡΠ°ΡΡΡΡ
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