Π Π°Π·ΡΠ°Π±ΠΎΡΠΊΠ° Π²Π΅ΠΊΡΠΎΡΠΎΠ² Π΄Π»Ρ ΠΌΠΎΠ»Π΅ΠΊΡΠ»ΡΡΠ½ΠΎΠ³ΠΎ ΠΊΠ»ΠΎΠ½ΠΈΡΠΎΠ²Π°Π½ΠΈΡ Ρ ΠΏΠΎΠ·ΠΈΡΠΈΠ²Π½ΠΎΠΉ ΡΠ΅Π»Π΅ΠΊΡΠΈΠ΅ΠΉ, ΠΎΡΠ½ΠΎΠ²Π°Π½Π½ΡΡ Π½Π° ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠΈ Π»Π΅ΡΠ°Π»ΡΠ½ΠΎΠ³ΠΎ ΡΡΡΠ΅ΠΊΡΠ° Π³Π΅Π½Π° Π±Π°ΡΠ½Π°Π·Ρ
ΠΠΏΠ΅ΡΠ²ΡΠ΅ ΡΠ°Π·ΡΠ°Π±ΠΎΡΠ°Π½Π° Π²ΡΡΠΎΠΊΠΎΡΡΡΠ΅ΠΊΡΠΈΠ²Π½Π°Ρ Π²Π΅ΠΊΡΠΎΡΠ½Π°Ρ ΡΠΈΡΡΠ΅ΠΌΠ° Π΄Π»Ρ ΠΌΠΎΠ»Π΅ΠΊΡΠ»ΡΡΠ½ΠΎΠ³ΠΎ ΠΊΠ»ΠΎΠ½ΠΈΡΠΎΠ²Π°Π½ΠΈΡ Ρ ΠΏΠΎΠ·ΠΈΡΠΈΠ²Π½ΠΎΠΉ ΡΠ΅Π»Π΅ΠΊΡΠΈΠ΅ΠΉ, ΠΈΡΠΏΠΎΠ»ΡΠ·ΡΡΡΠ°Ρ Π² ΠΊΠ°ΡΠ΅ΡΡΠ²Π΅ ΡΠ΅Π»Π΅ΠΊΡΠΈΠ²Π½ΠΎΠ³ΠΎ ΠΌΠ°ΡΠΊΠ΅ΡΠ° Π³Π΅Π½ ΡΠΈΠ±ΠΎΠ½ΡΠΊΠ»Π΅Π°Π·Ρ. ΠΠΎΠ»ΡΡΠ΅Π½Ρ Π²Π΅ΠΊΡΠΎΡΠ½ΡΠ΅ ΠΏΠ»Π°Π·ΠΌΠΈΠ΄Ρ, ΡΠΎΠ΄Π΅ΡΠΆΠ°ΡΠΈΠ΅ Π² ΠΊΠΎΠ΄ΠΈΡΡΡΡΠ΅ΠΉ ΡΠ°ΡΡΠΈ Π³Π΅Π½Π° ΡΠΈΠ±ΠΎΠ½ΡΠΊΠ»Π΅Π°Π·Ρ ΠΈΠ· Bacillus amyloliquefaciens — Π±Π°ΡΠ½Π°Π·Ρ ΡΡΠ°ΡΡΠΊΠΈ ΡΠ·Π½Π°Π²Π°Π½ΠΈΡ 2 (ΡΠΠ’438) ΠΈ 10 (ΡΠΠ’440) Π½Π°ΠΈΠ±ΠΎΠ»Π΅Π΅ ΡΠΏΠΎΡΡΠ΅Π±ΠΈΡΠ΅Π»ΡΠ½ΡΡ ΡΠ΅ΡΡΡΠΈΠΊΡΠΈΠΎΠ½Π½ΡΡ ΡΠ½Π΄ΠΎΠ½ΡΠΊΠ»Π΅Π°Π·. Π Π°Π·ΡΠ°Π±ΠΎΡΠ°Π½Π°… 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·ΠΎΠ²Π°Π½Π½ΡΡ ΡΠΎΠΊΡΠ°ΡΠ΅Π½ΠΈΠΉ
- ΠΠΠΠΠ 1. ΠΠΈΡΠ΅ΡΠ°ΡΡΡΠ½ΡΠΉ ΠΎΠ±Π·ΠΎΡ
- 1. 1. ΠΠ΅ΠΊΡΠΎΡΠ° Ρ ΠΏΠΎΠ·ΠΈΡΠΈΠ²Π½ΠΎΠΉ ΡΠ΅Π»Π΅ΠΊΡΠΈΠ΅ΠΉ
- 1. 2. Π ΠΠΠ°Π·Π° Π±Π°ΡΠ½Π°Π·Π° ΠΊΠ°ΠΊ ΡΠ°ΠΊΡΠΎΡ Π»Π΅ΡΠ°Π»ΡΠ½ΠΎΡΡΠΈ
- 1. 2. 1. Π₯Π°ΡΠ°ΠΊΡΠ΅ΡΠΈΡΡΠΈΠΊΠ° ΡΠ΅ΡΠΌΠ΅Π½ΡΠ°
- 1. 2. 2. ΠΠ½ΡΡΡΠΈΠΊΠ»Π΅ΡΠΎΡΠ½ΡΠΉ ΠΈΠ½Π³ΠΈΠ±ΠΈΡΠΎΡ Π±Π°ΡΠ½Π°Π·Ρ — Π±Π°ΡΡΡΠ°Ρ
- 3. 1. Π Π°Π·ΡΠ°Π±ΠΎΡΠΊΠ° Π½Π° ΠΎΡΠ½ΠΎΠ²Π΅ Π³Π΅Π½Π° Π±Π°ΡΠ½Π°Π·Ρ Π²Π΅ΠΊΡΠΎΡΠ° ΡΠΠ’440 Ρ ΠΏΠΎΠ·ΠΈΡΠΈΠ²Π½ΠΎΠΉ ΡΠ΅Π»Π΅ΠΊΡΠΈΠ΅ΠΉ
- 3. 1. 1. ΠΠ·ΡΡΠ΅Π½ΠΈΠ΅ Π²Π»ΠΈΡΠ½ΠΈΡ ΡΡΠΎΠ²Π½Ρ ΠΈΠ½Π΄ΡΠΊΡΠΈΠΈ tac—ΠΏΡΠΎΠΌΠΎΡΠΎΡΠ° Π½Π° Π²ΡΠΆΠΈΠ²Π°Π΅ΠΌΠΎΡΡΡ ΡΠ°Π·Π»ΠΈΡΠ½ΡΡ ΡΡΠ°ΠΌΠΌΠΎΠ² E. coli, ΡΡΠ°Π½ΡΡΠΎΡΠΌΠΈΡΠΎΠ²Π°Π½Π½ΡΡ ΠΏΠ»Π°Π·ΠΌΠΈΠ΄ΠΎΠΉ ΡΠΠ’
- 3. 1. 2. ΠΠ»ΠΎΠ½ΠΈΡΠΎΠ²Π°Π½ΠΈΠ΅ Ρ ΠΏΠΎΠΌΠΎΡΡΡ ΠΏΠ»Π°Π·ΠΌΠΈΠ΄Ρ ΡΠΠ’416 ΠΠ¦Π — ΠΏΡΠΎΠ΄ΡΠΊΡΠ°, ΠΊΠΎΠ΄ΠΈΡΡΡΡΠ΅Π³ΠΎ Π²Π°ΡΠΈΠ°Π±Π΅Π»ΡΠ½ΡΠ΅ ΡΠ΅Π³ΠΈΠΎΠ½Ρ Π»Π΅Π³ΠΊΠΎΠΉ ΡΠ΅ΠΏΠΈ Π°Π½ΡΠΈΡΠ΅Π»Π° ΠΊ Π±Π΅Π»ΠΊΡ Ρ24 Π²ΠΈΡΡΡΠ° HIV
- 3. 1. 3. ΠΠ²Π΅Π΄Π΅Π½ΠΈΠ΅ ΠΏΠΎΠ»ΠΈΠ»ΠΈΠ½ΠΊΠ΅ΡΠ° Π² Π³Π΅Π½ Π±Π°ΡΠ½Π°Π·Ρ ΠΈ ΠΊΠΎΠ½ΡΡΡΡΠΊΡΠΈΡ Π²Π΅ΠΊΡΠΎΡΠΎΠ² ΡΠΠ’438, ΡΠΠ’438Π° ΠΈ ΡΠΠ’
- 3. 1. 4. ΠΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠ΅ Π²Π΅ΠΊΡΠΎΡΠ° ΡΠΠ’440 Ρ ΠΏΠΎΠ·ΠΈΡΠΈΠ²Π½ΠΎΠΉ ΡΠ΅Π»Π΅ΠΊΡΠΈΠ΅ΠΉ Π΄Π»Ρ ΠΊΠ»ΠΎΠ½ΠΈΡΠΎΠ²Π°Π½ΠΈΡ ΡΡΠ°Π³ΠΌΠ΅Π½ΡΠΎΠ² ΡΡΠΆΠ΅ΡΠΎΠ΄Π½ΠΎΠΉ ΠΠΠ
- 3. 2. ΠΠΎΠ½ΡΡΡΡΠΈΡΠΎΠ²Π°Π½ΠΈΠ΅ Π²Π΅ΠΊΡΠΎΡΠ° ΡΠΠ° —7 Ρ ΠΏΠΎΠ·ΠΈΡΠΈΠ²Π½ΠΎΠΉ ΡΠ΅Π»Π΅ΠΊΡΠΈΠ΅ΠΉ.70 3.2.1. ΠΠΎΠ½ΡΡΡΡΠΈΡΠΎΠ²Π°Π½ΠΈΠ΅ ΠΏΠ»Π°Π·ΠΌΠΈΠ΄ ΡΠΠ° —2, ΡΠΠ° — 3s ΠΈ ΡΠΠ°
- 3. 2. 2. Π‘Π°ΠΉΡ — ΡΠΏΠ΅ΡΠΈΡΠΈΡΠ΅ΡΠΊΠΈΠΉ ΠΌΡΡΠ°Π³Π΅Π½Π΅Π· Π΄Π»Ρ ΡΠ΄Π°Π»Π΅Π½ΠΈΡ. ΠΏΠΎΠ²ΡΠΎΡΡΡΡΠΈΡ ΡΡ ΡΠ°ΠΉΡΠΎΠ² ΡΠ΅ΡΡΡΠΈΠΊΡΠΈΠΈ ΠΈ ΠΊΠΎΠ½ΡΡΡΡΠΊΡΠΈΡ Π²Π΅ΠΊΡΠΎΡΠ° ΡΠΠ°
- 3. 2. 3. ΠΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠ΅ Π²Π΅ΠΊΡΠΎΡΠ° ΡΠΠ° —7 Π΄Π»Ρ ΠΊΠ»ΠΎΠ½ΠΈΡΠΎΠ²Π°Π½ΠΈΡ ΡΡΠ°Π³ΠΌΠ΅Π½ΡΠΎΠ² ΡΡΠΆΠ΅ΡΠΎΠ΄Π½ΠΎΠΉ ΠΠΠ
- 3. 2. 4. ΠΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠ΅ Π²Π΅ΠΊΡΠΎΡΠ° ΡΠΠ° — 7 Π΄Π»Ρ ΠΊΠ»ΠΎΠ½ΠΈΡΠΎΠ²Π°Π½ΠΈΡ Π³Π΅Π½ΠΎΠ² ΠΈΠΌΠΌΡΠ½ΠΎΠ³Π»ΠΎΠ±ΡΠ»ΠΈΠ½ΠΎΠ² ΠΌΡΡΠΈ
- 3. 3. ΠΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΠ΅ Ρ ΠΏΠΎΠΌΠΎΡΡΡ ΠΈΠΌΠΌΡΠ½ΠΎΠ±Π»ΠΎΡΠΈΠ½Π³Π° ΡΠ΅ΠΊΠΎΠΌΠ±ΠΈΠ½Π°Π½ΡΠ½ΠΎΠΉ Π±Π°ΡΠ½Π°Π·Ρ ΠΈ Π΅Π΅ ΠΌΡΡΠ°Π½ΡΠΎΠ², ΠΏΡΠΎΠ΄ΡΡΠΈΡΡΠ΅ΠΌΡΡ Ρ ΠΏΠΎΠΌΠΎΡΡΡ ΠΏΠ»Π°Π·ΠΌΠΈΠ΄ ΡΠΠ’440 ΠΈ ΡΠΠ°
- 3. 4. Π₯Π°ΡΠ°ΠΊΡΠ΅ΡΠΈΡΡΠΈΠΊΠ° Π²ΠΊΠ»Π°Π΄Π° ΡΡΠ΄Π° ΠΌΡΡΠ°ΡΠΈΠΉ Π² ΡΠ΅ΡΠΌΠ΅Π½ΡΠ°ΡΠΈΠ²Π½ΡΡ Π°ΠΊΡΠΈΠ²Π½ΠΎΡΡΡ ΠΌΠΎΠ΄ΠΈΡΠΈΡΠΈΡΠΎΠ²Π°Π½Π½ΠΎΠΉ Π±Π°ΡΠ½Π°Π·Ρ
Π Π°Π·ΡΠ°Π±ΠΎΡΠΊΠ° Π²Π΅ΠΊΡΠΎΡΠΎΠ² Π΄Π»Ρ ΠΌΠΎΠ»Π΅ΠΊΡΠ»ΡΡΠ½ΠΎΠ³ΠΎ ΠΊΠ»ΠΎΠ½ΠΈΡΠΎΠ²Π°Π½ΠΈΡ Ρ ΠΏΠΎΠ·ΠΈΡΠΈΠ²Π½ΠΎΠΉ ΡΠ΅Π»Π΅ΠΊΡΠΈΠ΅ΠΉ, ΠΎΡΠ½ΠΎΠ²Π°Π½Π½ΡΡ Π½Π° ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠΈ Π»Π΅ΡΠ°Π»ΡΠ½ΠΎΠ³ΠΎ ΡΡΡΠ΅ΠΊΡΠ° Π³Π΅Π½Π° Π±Π°ΡΠ½Π°Π·Ρ (ΡΠ΅ΡΠ΅ΡΠ°Ρ, ΠΊΡΡΡΠΎΠ²Π°Ρ, Π΄ΠΈΠΏΠ»ΠΎΠΌ, ΠΊΠΎΠ½ΡΡΠΎΠ»ΡΠ½Π°Ρ)
ΠΠΎΠ·ΡΠ°ΡΡΠ°ΡΡΠ°Ρ ΠΈΠ½ΡΠ΅Π½ΡΠΈΠ²Π½ΠΎΡΡΡ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΠΉ ΠΏΠΎ ΠΌΠΎΠ»Π΅ΠΊΡΠ»ΡΡΠ½ΠΎΠΉ Π±ΠΈΠΎΠ»ΠΎΠ³ΠΈΠΈ ΠΈ Π³Π΅Π½Π΅ΡΠΈΠΊΠ΅ ΡΡΠ΅Π±ΡΠ΅Ρ ΠΏΠΎΠ²ΡΡΠ΅Π½ΠΈΡ ΡΡΡΠ΅ΠΊΡΠΈΠ²Π½ΠΎΡΡΠΈ ΠΎΠ΄Π½ΠΎΠΉ ΠΈΠ· ΠΎΡΠ½ΠΎΠ²Π½ΡΡ ΠΌΠ΅ΡΠΎΠ΄ΠΈΡΠ΅ΡΠΊΠΈΡ ΠΏΡΠΎΡΠ΅Π΄ΡΡ — ΠΌΠΎΠ»Π΅ΠΊΡΠ»ΡΡΠ½ΠΎΠ³ΠΎ ΠΊΠ»ΠΎΠ½ΠΈΡΠΎΠ²Π°Π½ΠΈΡ. Π ΠΏΠΎΡΠ»Π΅Π΄Π½ΠΈΠ΅ Π³ΠΎΠ΄Ρ Π±ΡΡΡΡΠΎ ΡΠ°Π·Π²ΠΈΠ²Π°Π΅ΡΡΡ Π½ΠΎΠ²ΠΎΠ΅ Π±ΠΈΠΎΡΠ΅Ρ Π½ΠΎΠ»ΠΎΠ³ΠΈΡΠ΅ΡΠΊΠΎΠ΅ Π½Π°ΠΏΡΠ°Π²Π»Π΅Π½ΠΈΠ΅ — Π²Π΅ΠΊΡΠΎΡΠ½ΡΠ΅ ΡΠΈΡΡΠ΅ΠΌΡ Ρ ΠΏΠΎΠ·ΠΈΡΠΈΠ²Π½ΠΎΠΉ ΡΠ΅Π»Π΅ΠΊΡΠΈΠ΅ΠΉ, ΠΏΡΠΈ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠΈ ΠΊΠΎΡΠΎΡΡΡ Π²ΡΠΆΠΈΠ²Π°ΡΡ ΡΠΎΠ»ΡΠΊΠΎ ΠΊΠ»ΠΎΠ½Ρ, ΡΠΎΠ΄Π΅ΡΠΆΠ°ΡΠΈΠ΅ ΡΠ΅ΠΊΠΎΠΌΠ±ΠΈΠ½Π°Π½ΡΠ½ΡΠ΅ ΠΏΠ»Π°Π·ΠΌΠΈΠ΄Ρ. ΠΡΠ΅Π²ΠΈΠ΄Π½Ρ ΠΏΡΠ΅ΠΈΠΌΡΡΠ΅ΡΡΠ²Π° ΡΠ°ΠΊΠΈΡ Π²Π΅ΠΊΡΠΎΡΠΎΠ², ΠΊΠΎΡΠΎΡΡΠ΅ Π½Π΅ ΡΡΠ΅Π±ΡΡΡ Π΄Π΅ΡΠΎΡΡΠΎΡΠΈΠ»ΠΈΡΠΎΠ²Π°Π½ΠΈΡ Π»ΠΈΠ½Π΅Π°ΡΠΈΠ·ΠΈΡΠΎΠ²Π°Π½Π½ΠΎΠΉ ΠΏΠ»Π°Π·ΠΌΠΈΠ΄Ρ Π΄Π»Ρ ΠΊΠ»ΠΎΠ½ΠΈΡΠΎΠ²Π°Π½ΠΈΡ. ΠΡΠΈ ΡΡΠΎΠΌ ΠΎΡΡΡΡΡΡΠ²ΡΠ΅Ρ Π²Π»ΠΈΡΠ½ΠΈΠ΅ Π½Π΅ΡΠ°Π·ΡΠ΅Π·Π°Π½Π½ΠΎΠ³ΠΎ Π²Π΅ΠΊΡΠΎΡΠ°, ΠΏΡΠΈΡΡΡΡΡΠ²ΡΡΡΠ΅Π³ΠΎ ΠΏΡΠ°ΠΊΡΠΈΡΠ΅ΡΠΊΠΈ Π² Π»ΡΠ±ΡΡ ΠΏΡΠ΅ΠΏΠ°ΡΠ°ΡΠ°Ρ . ΠΡΡΠ³ΠΈΠΌ ΠΏΡΠ΅ΠΈΠΌΡΡΠ΅ΡΡΠ²ΠΎΠΌ ΡΠ²Π»ΡΠ΅ΡΡΡ ΠΎΡΡΡΡΡΡΠ²ΠΈΠ΅ Π½Π΅ΠΎΠ±Ρ ΠΎΠ΄ΠΈΠΌΠΎΡΡΠΈ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΡ Π΄ΠΎΡΠΎΠ³ΠΎΡΡΠΎΡΡΠΈΡ ΡΠ΅Π°Π³Π΅Π½ΡΠΎΠ², ΡΠ°ΠΊΠΈΡ ΠΊΠ°ΠΊ X —gal, Π΄ΠΎΠΏΠΎΠ»Π½ΠΈΡΠ΅Π»ΡΠ½ΡΠ΅ Π°Π½ΡΠΈΠ±ΠΈΠΎΡΠΈΠΊΠΈ ΠΈ ΠΏΡ. ΠΡΠ΅ ΠΏΠ΅ΡΠ΅ΡΠΈΡΠ»Π΅Π½Π½ΠΎΠ΅ Π²ΡΡΠ΅ ΡΠ½ΠΈΠΆΠ°Π΅Ρ ΡΡΠΎΠΈΠΌΠΎΡΡΡ ΠΈ ΠΏΠΎΠ²ΡΡΠ°Π΅Ρ ΡΡΡΠ΅ΠΊΡΠΈΠ²Π½ΠΎΡΡΡ Π³Π΅Π½Π½ΠΎ — ΠΈΠ½ΠΆΠ΅Π½Π΅ΡΠ½ΡΡ ΠΌΠ°Π½ΠΈΠΏΡΠ»ΡΡΠΈΠΉ. ΠΠ΅ΡΠ²ΡΠ΅ Π²Π΅ΠΊΡΠΎΡΠ° Π½ΠΎΠ²ΠΎΠ³ΠΎ ΠΏΠΎΠΊΠΎΠ»Π΅Π½ΠΈΡ ΡΠΆΠ΅ ΠΏΠΎΡΠ²ΠΈΠ»ΠΈΡΡ Π² ΠΏΡΠΎΠ΄Π°ΠΆΠ΅, Π½Π°ΠΏΡΠΈΠΌΠ΅Ρ Π²Π΅ΠΊΡΠΎΡ pZErO-1™ (Invitrogen Inc., USA), ΠΎΡΠ½ΠΎΠ²Π°Π½Π½ΡΠΉ Π½Π° ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠΈ Π»Π΅ΡΠ°Π»ΡΠ½ΠΎΠ³ΠΎ ΡΡΡΠ΅ΠΊΡΠ° Π³Π΅Π½Π° ccdB (control of cell death Π gene) [1]. ΠΠ΄Π½Π°ΠΊΠΎ Π² ΡΡΠ΄Π΅ ΡΠ»ΡΡΠ°Π΅Π² Π³Π΅Π½Ρ, ΠΎΠΏΡΠ΅Π΄Π΅Π»ΡΡΡΠΈΠ΅ Π»Π΅ΡΠ°Π»ΡΠ½ΠΎΡΡΡ, ΡΠ»ΠΈΡΠΊΠΎΠΌ Π²Π΅Π»ΠΈΠΊΠΈ, ΡΡΠΎ ΠΏΡΠΈΠ²ΠΎΠ΄ΠΈΡ ΠΊ Π·Π½Π°ΡΠΈΡΠ΅Π»ΡΠ½ΠΎΠΌΡ ΠΏΡΠ΅Π²ΡΡΠ΅Π½ΠΈΡ ΠΎΠΏΡΠΈΠΌΠ°Π»ΡΠ½ΡΡ ΡΠ°Π·ΠΌΠ΅ΡΠΎΠ² ΠΏΠ»Π°Π·ΠΌΠΈΠ΄Ρ. ΠΠΈΠ±ΠΎ ΠΈΠ·Π²Π΅ΡΡΠ½ΡΠ΅ Π²Π΅ΠΊΡΠΎΡΠ° Ρ ΠΏΠΎΠ·ΠΈΡΠΈΠ²Π½ΠΎΠΉ ΡΠ΅Π»Π΅ΠΊΡΠΈΠ΅ΠΉ ΡΡΠ΅Π±ΡΡΡ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΡ ΡΠΏΠ΅ΡΠΈΠ°Π»ΡΠ½ΡΡ ΡΡΠ°ΠΌΠΌΠΎΠ² E. coli ΠΈΠ»ΠΈ Π΄ΠΎΠΏΠΎΠ»Π½ΠΈΡΠ΅Π»ΡΠ½ΡΡ ΠΊΠΎΠΌΠΏΠΎΠ½Π΅Π½ΡΠΎΠ² ΠΏΠΈΡΠ°ΡΠ΅Π»ΡΠ½ΡΡ ΡΡΠ΅Π΄.
ΠΠΎΠ»ΡΡΠΈΠ½ΡΡΠ²ΠΎ ΠΎΠΏΠΈΡΠ°Π½Π½ΡΡ Π² Π»ΠΈΡΠ΅ΡΠ°ΡΡΡΠ΅ Π²Π΅ΠΊΡΠΎΡΠΎΠ² Ρ ΠΏΠΎΠ·ΠΈΡΠΈΠ²Π½ΠΎΠΉ ΡΠ΅Π»Π΅ΠΊΡΠΈΠ΅ΠΉ Π½Π΅ ΠΎΠ±Π»Π°Π΄Π°ΡΡ Π²ΡΠ΅ΠΌΠΈ Π½Π΅ΠΎΠ±Ρ ΠΎΠ΄ΠΈΠΌΡΠΌΠΈ Π°ΡΡΠΈΠ±ΡΡΠ°ΠΌΠΈ ΡΠΎΠ²ΡΠ΅ΠΌΠ΅Π½Π½ΡΡ Π²Π΅ΠΊΡΠΎΡΠΎΠ², ΡΠ°ΠΊΠΈΠΌΠΈ, ΠΊΠ°ΠΊ ΡΠ΅ΠΊΠ²Π΅Π½ΠΈΡΠΎΠ²Π°Π½ΠΈΠ΅ Ρ ΠΏΠΎΠΌΠΎΡΡΡ ΡΠ½ΠΈΠ²Π΅ΡΡΠ°Π»ΡΠ½ΡΡ ΠΏΡΠ°ΠΉΠΌΠ΅ΡΠΎΠ², ΠΊΠ»ΠΎΠ½ΠΈΡΠΎΠ²Π°Π½ΠΈΠ΅ ΠΏΡΠΎΡΡΠΆΠ΅Π½Π½ΡΡ ΡΡΠ°Π³ΠΌΠ΅Π½ΡΠΎΠ² ΡΡΠΆΠ΅ΡΠΎΠ΄Π½ΠΎΠΉ ΠΠΠ Ρ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠ΅ΠΌ ΡΠΈΡΠΎΠΊΠΎΠ³ΠΎ ΡΠΏΠ΅ΠΊΡΡΠ° ΡΠ΅ΡΡΡΠΈΠΊΡΠ°Π·, ΠΏΠΎΠ»ΡΡΠ΅Π½ΠΈΠ΅ ΠΎΠ΄Π½ΠΎΡΠ΅ΠΏΠΎΡΠ΅ΡΠ½ΠΎΠΉ ΠΠΠ, in vitro ΡΡΠ°Π½ΡΠΊΡΠΈΠΏΡΠΈΡ. ΠΡΠ΅Π΄ΡΡΠ°Π²Π»ΡΠ΅ΡΡΡ Π°ΠΊΡΡΠ°Π»ΡΠ½ΠΎΠΉ Π΄Π°Π»ΡΠ½Π΅ΠΉΡΠ°Ρ ΡΠ°Π·ΡΠ°Π±ΠΎΡΠΊΠ° ΡΠΎΠ²ΡΠ΅ΠΌΠ΅Π½Π½ΡΡ Π²Π΅ΠΊΡΠΎΡΠΎΠ² Ρ ΠΏΠΎΠ·ΠΈΡΠΈΠ²Π½ΠΎΠΉ ΡΠ΅Π»Π΅ΠΊΡΠΈΠ΅ΠΉ, Π½Π΅ ΠΈΠΌΠ΅ΡΡΠΈΡ Π²ΡΡΠ΅ΠΏΠ΅ΡΠ΅ΡΠΈΡΠ»Π΅Π½Π½ΡΡ Π½Π΅Π΄ΠΎΡΡΠ°ΡΠΊΠΎΠ². Π Π½Π°ΡΡΠΎΡΡΠ΅ΠΉ ΡΠ°Π±ΠΎΡΠ΅ Π²ΠΏΠ΅ΡΠ²ΡΠ΅ ΠΏΠΎΠ»ΡΡΠ΅Π½Ρ Π²Π΅ΠΊΡΠΎΡΠ° Ρ ΠΏΠΎΠ·ΠΈΡΠΈΠ²Π½ΠΎΠΉ ΡΠ΅Π»Π΅ΠΊΡΠΈΠ΅ΠΉ, ΠΎΡΠ½ΠΎΠ²Π°Π½Π½ΡΠ΅ Π½Π° ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠΈ Π»Π΅ΡΠ°Π»ΡΠ½ΠΎΠ³ΠΎ ΡΡΡΠ΅ΠΊΡΠ° Π³Π΅Π½Π° Π±Π°ΡΠ½Π°Π·Ρ — ΡΠΈΠ±ΠΎΠ½ΡΠΊΠ»Π΅Π°Π·Ρ ΠΈΠ· Bacillus amyloliquefaciens, Π²ΡΠ·ΡΠ²Π°ΡΡΠ΅ΠΉ ΠΏΡΠΈ ΡΠΊΡΠΏΡΠ΅ΡΡΠΈΠΈ Π² Π±Π°ΠΊΡΠ΅ΡΠΈΠ°Π»ΡΠ½ΡΡ ΠΊΠ»Π΅ΡΠΊΠ°Ρ Π»Π΅ΡΠ°Π»ΡΠ½ΡΠΉ Π³ΠΈΠ΄ΡΠΎΠ»ΠΈΠ· Π²Π½ΡΡΡΠΈΠΊΠ»Π΅ΡΠΎΡΠ½ΠΎΠΉ Π ΠΠ [2,3]. ΠΠΎΠ»ΡΡΠ΅Π½Π½ΡΠ΅ Π²Π΅ΠΊΡΠΎΡΠ½ΡΠ΅ ΡΠΈΡΡΠ΅ΠΌΡ ΡΡΠ½ΠΊΡΠΈΠΎΠ½ΠΈΡΡΡΡ ΡΠ°ΠΊΠΈΠΌ ΠΎΠ±ΡΠ°Π·ΠΎΠΌ, ΡΡΠΎ ΠΊΠ»ΠΎΠ½ΠΈΡΠΎΠ²Π°Π½ΠΈΠ΅ ΡΡΠ°Π³ΠΌΠ΅Π½ΡΠΎΠ² ΠΠΠ Π² ΠΏΠΎΠ»ΠΈΠ»ΠΈΠ½ΠΊΠ΅ΡΡ ΡΡΠΈΡ Π²Π΅ΠΊΡΠΎΡΠΎΠ² ΠΏΡΠΈΠ²ΠΎΠ΄ΠΈΡ ΠΊ ΡΡΡΡΠΊΡΡΡΠ½ΠΎΠΉ ΠΈΠ½Π°ΠΊΡΠΈΠ²Π°ΡΠΈΠΈ Π³Π΅Π½Π° Π±Π°ΡΠ½Π°Π·Ρ, Π² ΡΠ΅Π·ΡΠ»ΡΡΠ°ΡΠ΅ ΡΠ΅Π³ΠΎ Π²ΡΡΠ°ΡΡΠ°ΡΡ ΡΠΎΠ»ΡΠΊΠΎ ΠΊΠ»ΠΎΠ½Ρ ΡΡΠ°Π½ΡΡΠΎΡΠΌΠ°Π½ΡΠΎΠ², ΡΠΎΠ΄Π΅ΡΠΆΠ°ΡΠΈΡ Π²ΡΡΠ°Π²ΠΊΠΈ ΡΡΠΆΠ΅ΡΠΎΠ΄Π½ΠΎΠΉ ΠΠΠ. Π’ΡΠ°Π½ΡΡΠΎΡΠΌΠ°Π½ΡΡ, ΡΠΎΠ΄Π΅ΡΠΆΠ°ΡΠΈΠ΅ ΡΠ΅Π»ΠΈΠ³ΠΈΡΠΎΠ²Π°Π½Π½ΡΡ ΠΏΠ»Π°Π·ΠΌΠΈΠ΄Ρ, ΠΏΠΎΠ³ΠΈΠ±Π°ΡΡ ΠΏΡΠΈ Π²ΡΡΠ°ΡΠΈΠ²Π°Π½ΠΈΠΈ Π² ΠΏΡΠΈΡΡΡΡΡΠ²ΠΈΠΈ ΠΠΠ’Π, ΠΈΠ½Π΄ΡΡΠΈΡΡΡΡΠ΅Π³ΠΎ ΡΠΊΡΠΏΡΠ΅ΡΡΠΈΡ Π±Π°ΡΠ½Π°Π·Ρ Π² ΠΊΠ»Π΅ΡΠΊΠ°Ρ E.coli. ΠΠΎΠ»ΡΡΠ΅Π½Π½ΡΠ΅ Π²Π΅ΠΊΡΠΎΡΠ° ΡΠΠ’440 ΠΈ ΡΠ°Π³Π΅ΠΌΠΈΠ΄Π° ΡΠΠ° —7 ΠΌΠΎΠ³ΡΡ Π±ΡΡΡ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½Ρ Π² ΡΠΈΡΠΎΠΊΠΎΠΌ ΡΠΏΠ΅ΠΊΡΡΠ΅ ΡΡΠ°ΠΌΠΌΠΎΠ² E. coli ΠΈ ΠΏΠΎΠ·Π²ΠΎΠ»ΡΡΡ Π²ΡΠΏΠΎΠ»Π½ΡΡΡ ΠΏΠΎΠ»Π½ΡΠΉ ΡΠΏΠ΅ΠΊΡΡ Π³Π΅Π½Π½ΠΎ-ΠΈΠ½ΠΆΠ΅Π½Π΅ΡΠ½ΡΡ ΠΌΠ°Π½ΠΈΠΏΡΠ»ΡΡΠΈΠΉ, Ρ Π°ΡΠ°ΠΊΡΠ΅ΡΠ½ΡΠΉ Π΄Π»Ρ ΡΠΎΠ²ΡΠ΅ΠΌΠ΅Π½Π½ΡΡ Π²Π΅ΠΊΡΠΎΡΠΎΠ².
Π¦Π΅Π»ΡΡ Π½Π°ΡΡΠΎΡΡΠ΅Π³ΠΎ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΡ ΡΠ²Π»ΡΠ»Π°ΡΡ ΡΠ°Π·ΡΠ°Π±ΠΎΡΠΊΠ° Π½ΠΎΠ²ΡΡ Π²Π΅ΠΊΡΠΎΡΠ½ΡΡ ΡΠΈΡΡΠ΅ΠΌ Ρ ΠΏΠΎΠ·ΠΈΡΠΈΠ²Π½ΠΎΠΉ ΡΠ΅Π»Π΅ΠΊΡΠΈΠ΅ΠΉ Π΄Π»Ρ ΠΊΠ»ΠΎΠ½ΠΈΡΠΎΠ²Π°Π½ΠΈΡ ΡΡΠ°Π³ΠΌΠ΅Π½ΡΠΎΠ² ΡΡΠΆΠ΅ΡΠΎΠ΄Π½ΠΎΠΉ ΠΠΠ, ΠΈΡΠΏΠΎΠ»ΡΠ·ΡΡΡΠΈΡ Π»Π΅ΡΠ°Π»ΡΠ½ΡΠΉ ΡΡΡΠ΅ΠΊΡ Π³Π΅Π½Π° Π±Π°ΡΠ½Π°Π·Ρ. ΠΡΡ ΠΎΠ΄Ρ ΠΈΠ· ΠΏΠΎΡΡΠ°Π²Π»Π΅Π½Π½ΠΎΠΉ ΡΠ΅Π»ΠΈ, Π±ΡΠ»ΠΈ ΡΡΠΎΡΠΌΡΠ»ΠΈΡΠΎΠ²Π°Π½Ρ ΡΠ»Π΅Π΄ΡΡΡΠΈΠ΅ Π·Π°Π΄Π°ΡΠΈ: Π°) ΠΈΠ·ΡΡΠΈΡΡ ΠΏΠ»Π°Π·ΠΌΠΈΠ΄Ρ ΡΠ416, ΠΏΡΠ΅Π΄ΡΡΠ°Π²Π»ΡΡΡΡΡ ΡΠΎΠ±ΠΎΠΉ Π²Π΅ΠΊΡΠΎΡ Π΄Π»Ρ ΡΠΊΡΠΏΡΠ΅ΡΡΠΈΠΈ Π±Π°ΡΠ½Π°Π·Ρ Π² ΠΊΠ»Π΅ΡΠΊΠ°Ρ E. coli, ΡΠΎΠ΄Π΅ΡΠΆΠ°ΡΠΈΠΉ ΠΊΠ°ΡΡΠ΅ΡΡ Π³Π΅Π½ΠΎΠ² Π±Π°ΡΠ½Π°Π·Π° —Π±Π°ΡΡΡΠ°Ρ, Π½Π° ΠΏΡΠ΅Π΄ΠΌΠ΅Ρ Π»Π΅ΡΠ°Π»ΡΠ½ΠΎΡΡΠΈ Π΄Π»Ρ ΡΠ°Π·Π»ΠΈΡΠ½ΡΡ ΡΡΠ°ΠΌΠΌΠΎΠ² E. coliΠ±) ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°ΡΡ ΠΏΠ»Π°Π·ΠΌΠΈΠ΄Ρ ΡΠΠ’416 Π² ΠΊΠ°ΡΠ΅ΡΡΠ²Π΅ ΠΌΠΎΠ΄Π΅Π»ΡΠ½ΠΎΠ³ΠΎ Π²Π΅ΠΊΡΠΎΡΠ° Ρ ΠΏΠΎΠ·ΠΈΡΠΈΠ²Π½ΠΎΠΉ ΡΠ΅Π»Π΅ΠΊΡΠΈΠ΅ΠΉ Π΄Π»Ρ ΠΊΠ»ΠΎΠ½ΠΈΡΠΎΠ²Π°Π½ΠΈΡ ΡΡΠ°Π³ΠΌΠ΅Π½ΡΠΎΠ² ΡΡΠΆΠ΅ΡΠΎΠ΄Π½ΠΎΠΉ ΠΠΠΠ²) ΡΠ°Π·ΡΠ°Π±ΠΎΡΠ°ΡΡ Π½Π° ΠΎΡΠ½ΠΎΠ²Π΅ ΠΏΠ»Π°Π·ΠΌΠΈΠ΄Ρ ΡΠΠ’416 Π²Π΅ΠΊΡΠΎΡ Ρ ΠΏΠΎΠ·ΠΈΡΠΈΠ²Π½ΠΎΠΉ ΡΠ΅Π»Π΅ΠΊΡΠΈΠ΅ΠΉ, ΡΠΎΠ΄Π΅ΡΠΆΠ°ΡΠΈΠΉ ΠΏΡΠΎΡΡΠΆΠ΅Π½Π½ΡΠΉ ΠΏΠΎΠ»ΠΈΠ»ΠΈΠ½ΠΊΠ΅Ρ ΠΈΠ· pUC19 Π²Π½ΡΡΡΠΈ Π³Π΅Π½Π° Π±Π°ΡΠ½Π°Π·ΡΠ³) ΠΈΠ·ΡΡΠΈΡΡ ΡΡΡΠ΅ΠΊΡΠΈΠ²Π½ΠΎΡΡΡ Π½ΠΎΠ²ΠΎΠΉ Π²Π΅ΠΊΡΠΎΡΠ½ΠΎΠΉ ΡΠΈΡΡΠ΅ΠΌΡ Ρ ΠΏΠΎΠ·ΠΈΡΠΈΠ²Π½ΠΎΠΉ ΡΠ΅Π»Π΅ΠΊΡΠΈΠ΅ΠΉ Π΄Π»Ρ ΠΊΠ»ΠΎΠ½ΠΈΡΠΎΠ²Π°Π½ΠΈΡ ΡΡΠ°Π³ΠΌΠ΅Π½ΡΠΎΠ² ΡΡΠΆΠ΅ΡΠΎΠ΄Π½ΠΎΠΉ ΠΠΠΠ΄) ΡΠ°Π·ΡΠ°Π±ΠΎΡΠ°ΡΡ Π½Π° ΠΎΡΠ½ΠΎΠ²Π΅ Π²Π΅ΠΊΡΠΎΡΠ° pBluescript KS II + (Stratagene, USA) ΡΠΎΠ²ΡΠ΅ΠΌΠ΅Π½Π½ΡΡ ΠΏΠΎΠ»ΠΈΡΡΠ½ΠΊΡΠΈΠΎΠ½Π°Π»ΡΠ½ΡΡ Π²Π΅ΠΊΡΠΎΡΠ½ΡΡ ΡΠΈΡΡΠ΅ΠΌΡ Ρ ΠΏΠΎΠ·ΠΈΡΠΈΠ²Π½ΠΎΠΉ ΡΠ΅Π»Π΅ΠΊΡΠΈΠ΅ΠΉ, ΠΎΠ±Π»Π°Π΄Π°ΡΡΡΡ ΡΡΠ΄ΠΎΠΌ ΡΠΏΠ΅ΡΠΈΡΠΈΡΠ΅ΡΠΊΠΈΡ ΡΡΠ½ΠΊΡΠΈΠΉ, ΡΠ°ΠΊΠΈΡ , ΠΊΠ°ΠΊ ΡΠΈΠ½ΡΠ΅Π· Π ΠΠ in vitro (ΠΏΡΠΎΠΌΠΎΡΠΎΡ Π’Π Π ΠΠ — ΠΏΠΎΠ»ΠΈΠΌΠ΅ΡΠ°Π·Ρ), ΠΏΠΎΠ»ΡΡΠ΅Π½ΠΈΠ΅ ΠΎΠ΄Π½ΠΎΡΠ΅ΠΏΠΎΡΠ΅ΡΠ½ΠΎΠΉ ΠΠΠ (fl ΠΎΡΠΈΠ΄ΠΆΠΈΠ½), ΡΠ΅ΠΊΠ²Π΅Π½ΠΈΡΠΎΠ²Π°Π½ΠΈΠ΅ Ρ ΠΏΠΎΠΌΠΎΡΡΡ ΡΠ½ΠΈΠ²Π΅ΡΠ°Π»ΡΠ½ΡΡ ΠΏΡΠ°ΠΉΠΌΠ΅ΡΠΎΠ² (Π13 rev, Π’Π, KS, SK), ΠΊΠ»ΠΎΠ½ΠΈΡΠΎΠ²Π°Π½ΠΈΠ΅ ΠΏΡΠΎΡΡΠΆΠ΅Π½Π½ΡΡ ΡΡΠ°Π³ΠΌΠ΅Π½ΡΠΎΠ² ΡΡΠΆΠ΅ΡΠΎΠ΄Π½ΠΎΠΉ ΠΠΠ Ρ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠ΅ΠΌ ΡΠΈΡΠΎΠΊΠΎΠ³ΠΎ ΡΠΏΠ΅ΠΊΡΡΠ° ΡΠ΅ΡΡΡΠΈΠΊΡΠ°Π·.
ΠΠΠΠΠ 4. ΠΠ«ΠΠΠΠ«.
1. ΠΠΏΠ΅ΡΠ²ΡΠ΅ ΡΠ°Π·ΡΠ°Π±ΠΎΡΠ°Π½Π° Π²ΡΡΠΎΠΊΠΎΡΡΡΠ΅ΠΊΡΠΈΠ²Π½Π°Ρ Π²Π΅ΠΊΡΠΎΡΠ½Π°Ρ ΡΠΈΡΡΠ΅ΠΌΠ° Π΄Π»Ρ ΠΌΠΎΠ»Π΅ΠΊΡΠ»ΡΡΠ½ΠΎΠ³ΠΎ ΠΊΠ»ΠΎΠ½ΠΈΡΠΎΠ²Π°Π½ΠΈΡ Ρ ΠΏΠΎΠ·ΠΈΡΠΈΠ²Π½ΠΎΠΉ ΡΠ΅Π»Π΅ΠΊΡΠΈΠ΅ΠΉ, ΠΈΡΠΏΠΎΠ»ΡΠ·ΡΡΡΠ°Ρ Π² ΠΊΠ°ΡΠ΅ΡΡΠ²Π΅ ΡΠ΅Π»Π΅ΠΊΡΠΈΠ²Π½ΠΎΠ³ΠΎ ΠΌΠ°ΡΠΊΠ΅ΡΠ° Π³Π΅Π½ ΡΠΈΠ±ΠΎΠ½ΡΠΊΠ»Π΅Π°Π·Ρ. ΠΠΎΠ»ΡΡΠ΅Π½Ρ Π²Π΅ΠΊΡΠΎΡΠ½ΡΠ΅ ΠΏΠ»Π°Π·ΠΌΠΈΠ΄Ρ, ΡΠΎΠ΄Π΅ΡΠΆΠ°ΡΠΈΠ΅ Π² ΠΊΠΎΠ΄ΠΈΡΡΡΡΠ΅ΠΉ ΡΠ°ΡΡΠΈ Π³Π΅Π½Π° ΡΠΈΠ±ΠΎΠ½ΡΠΊΠ»Π΅Π°Π·Ρ ΠΈΠ· Bacillus amyloliquefaciens — Π±Π°ΡΠ½Π°Π·Ρ ΡΡΠ°ΡΡΠΊΠΈ ΡΠ·Π½Π°Π²Π°Π½ΠΈΡ 2 (ΡΠΠ’438) ΠΈ 10 (ΡΠΠ’440) Π½Π°ΠΈΠ±ΠΎΠ»Π΅Π΅ ΡΠΏΠΎΡΡΠ΅Π±ΠΈΡΠ΅Π»ΡΠ½ΡΡ ΡΠ΅ΡΡΡΠΈΠΊΡΠΈΠΎΠ½Π½ΡΡ ΡΠ½Π΄ΠΎΠ½ΡΠΊΠ»Π΅Π°Π·.
2. ΠΠΏΠ΅ΡΠ²ΡΠ΅ ΠΏΠΎΠ»ΡΡΠ΅Π½Ρ ΠΈΠ½ΡΠ΅ΡΡΠΈΠΎΠ½Π½ΡΠ΅ ΠΌΡΡΠ°Π½ΡΡ Π±Π°ΡΠ½Π°Π·Ρ, ΡΠΎΠ΄Π΅ΡΠΆΠ°ΡΠΈΠ΅ Π²ΠΌΠ΅ΡΡΠΎ Val —36 Π²ΡΡΠ°Π²ΠΊΠΈ ΠΈΠ· 6 ΠΈ 19 Π°ΠΌΠΈΠ½ΠΎΠΊΠΈΡΠ»ΠΎΡ. ΠΡΠΎΠ΄Π΅ΠΌΠΎΠ½ΡΡΡΠΈΡΠΎΠ²Π°Π½ΠΎ Π½Π°Π»ΠΈΡΠΈΠ΅ ΡΠ΅ΡΠΌΠ΅Π½ΡΠ°ΡΠΈΠ²Π½ΠΎΠΉ Π°ΠΊΡΠΈΠ²Π½ΠΎΡΡΠΈ Ρ ΡΠ°ΠΊΠΈΡ ΠΌΡΡΠ°Π½ΡΠΎΠ², ΡΡΠΎ ΡΠΎΠ³Π»Π°ΡΡΠ΅ΡΡΡ Ρ Π΄Π°Π½Π½ΡΠΌΠΈ ΠΎ ΡΡΡΡΠΊΡΡΡΠ½ΠΎ — ΡΡΠ½ΠΊΡΠΈΠΎΠ½Π°Π»ΡΠ½ΠΎΠΉ ΠΎΡΠ³Π°Π½ΠΈΠ·Π°ΡΠΈΠΈ Π±Π°ΡΠ½Π°Π·Ρ, ΠΏΠΎΠ»ΠΎΠΆΠ΅Π½Π½ΡΠΌΠΈ Π² ΠΎΡΠ½ΠΎΠ²Ρ ΡΠ°Π·ΡΠ°Π±ΠΎΡΠ°Π½Π½ΠΎΠ³ΠΎ ΠΏΠΎΠ΄Ρ ΠΎΠ΄Π°.
3. ΠΠΏΠ΅ΡΠ²ΡΠ΅ ΡΠΊΠΎΠ½ΡΡΡΡΠΈΡΠΎΠ²Π°Π½ ΡΠ°Π³Π΅ΠΌΠΈΠ΄Π½ΡΠΉ Π²Π΅ΠΊΡΠΎΡ ΡΠΠ° —7 Ρ ΠΏΠΎΠ·ΠΈΡΠΈΠ²Π½ΠΎΠΉ ΡΠ΅Π»Π΅ΠΊΡΠΈΠ΅ΠΉ, ΠΈΡΠΏΠΎΠ»ΡΠ·ΡΡΡΠΈΠΉ Π»Π΅ΡΠ°Π»ΡΠ½ΡΠΉ ΡΡΡΠ΅ΠΊΡ Π³Π΅Π½Π° Π±Π°ΡΠ½Π°Π·Ρ. ΠΠ΅ΠΊΡΠΎΡ ΠΏΡΠ΅Π΄ΠΎΡΡΠ°Π²Π»ΡΠ΅Ρ Π²ΠΎΠ·ΠΌΠΎΠΆΠ½ΠΎΡΡΠΈ Π΄Π»Ρ ΠΊΠ»ΠΎΠ½ΠΈΡΠΎΠ²Π°Π½ΠΈΡ ΠΏΡΠΎΡΡΠΆΠ΅Π½Π½ΡΡ ΡΡΠ°Π³ΠΌΠ΅Π½ΡΠΎΠ² ΡΡΠΆΠ΅ΡΠΎΠ΄Π½ΠΎΠΉ ΠΠΠ Ρ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠ΅ΠΌ ΡΠΈΡΠΎΠΊΠΎΠ³ΠΎ Π½Π°Π±ΠΎΡΠ° ΡΠ΅ΡΡΡΠΈΠΊΡΠ°Π·, ΡΠ΅ΠΊΠ²Π΅Π½ΠΈΡΠΎΠ²Π°Π½ΠΈΡ Ρ ΠΏΠΎΠΌΠΎΡΡΡ ΡΠ½ΠΈΠ²Π΅ΡΡΠ°Π»ΡΠ½ΡΡ ΠΏΡΠ°ΠΉΠΌΠ΅ΡΠΎΠ², ΠΏΠΎΠ»ΡΡΠ΅Π½ΠΈΡ ΠΎΠ΄Π½ΠΎΡΠ΅ΠΏΠΎΡΠ΅ΡΠ½ΠΎΠΉ ΠΠΠ, Π° ΡΠ°ΠΊΠΆΠ΅ Π΄Π»Ρ ΡΠΈΠ½ΡΠ΅Π·Π° Π ΠΠ in vitro.
4. Π Π°Π·ΡΠ°Π±ΠΎΡΠ°Π½Π° ΠΏΠΎΠ»ΡΠΊΠΎΠ»ΠΈΡΠ΅ΡΡΠ²Π΅Π½Π½Π°Ρ ΠΌΠ΅ΡΠΎΠ΄ΠΈΠΊΠ° ΠΎΡΠ΅Π½ΠΊΠΈ ΡΠ΅ΡΠΌΠ΅Π½ΡΠ°ΡΠΈΠ²Π½ΠΎΠΉ Π°ΠΊΡΠΈΠ²Π½ΠΎΡΡΠΈ ΠΌΡΡΠ°Π½ΡΠΎΠ² Π±Π°ΡΠ½Π°Π·Ρ ΠΏΠΎ ΠΈΡ ΡΠΈΡΠΎΡΠΎΠΊΡΠΈΡΠ½ΠΎΡΡΠΈ, ΠΎΡΠ½ΠΎΠ²Π°Π½Π½Π°Ρ Π½Π° ΠΈΠ·ΠΌΠ΅ΡΠ΅Π½ΠΈΠΈ ΡΡΠ΅Π΄Π½Π΅ΡΡΠ°ΡΠΈΡΡΠΈΡΠ΅ΡΠΊΠΎΠ³ΠΎ Π΄ΠΈΠ°ΠΌΠ΅ΡΡΠ° Π±Π°ΠΊΡΠ΅ΡΠΈΠ°Π»ΡΠ½ΡΡ ΠΊΠΎΠ»ΠΎΠ½ΠΈΠΉ. Π‘ Π΅Π΅ ΠΏΠΎΠΌΠΎΡΡΡ ΠΎΡΠ΅Π½Π΅Π½ΠΎ Π²Π»ΠΈΡΠ½ΠΈΠ΅ ΠΠ¦Π — ΠΈΠ½Π΄ΡΡΠΈΡΠΎΠ²Π°Π½Π½ΡΡ ΠΌΡΡΠ°ΡΠΈΠΉ Π½Π° ΡΠ΅ΡΠΌΠ΅Π½ΡΠ°ΡΠΈΠ²Π½ΡΡ Π°ΠΊΡΠΈΠ²Π½ΠΎΡΡΡ ΠΌΠΎΠ΄ΠΈΡΠΈΡΠΈΡΠΎΠ²Π°Π½Π½ΠΎΠΉ Π±Π°ΡΠ½Π°Π·Ρ, ΡΠΎΠ΄Π΅ΡΠΆΠ°ΡΠ΅ΠΉ Π²ΡΡΠ°Π²ΠΊΡ ΠΈΠ· 6 Π°ΠΌΠΈΠ½ΠΎΠΊΠΈΡΠ»ΠΎΡ ΠΏΠΎ ΠΏΠΎΠ»ΠΎΠΆΠ΅Π½ΠΈΡ Val — 36.
5. ΠΡΡΠ΅ΠΊΡΠΈΠ²Π½ΠΎΡΡΡ ΡΠ°Π·ΡΠ°Π±ΠΎΡΠ°Π½Π½ΡΡ Π²Π΅ΠΊΡΠΎΡΠΎΠ² ΠΎΡΠ΅Π½Π΅Π½Π° ΠΊΠ°ΠΊ Π½Π° ΠΏΡΠΈΠΌΠ΅ΡΠ΅ ΠΊΠ»ΠΎΠ½ΠΈΡΠΎΠ²Π°Π½ΠΈΡ ΠΈΠ½Π΄ΠΈΠ²ΠΈΠ΄ΡΠ°Π»ΡΠ½ΡΡ ΡΡΠ°Π³ΠΌΠ΅Π½ΡΠΎΠ² ΠΠΠ, ΡΠ°ΠΊ ΠΈ Π½Π° ΠΏΡΠΈΠΌΠ΅ΡΠ°Ρ ΠΏΠΎΠ»ΡΡΠ΅Π½ΠΈΡ Π³Π΅Π½ΠΎΠΌΠ½ΠΎΠΉ Π±ΠΈΠ±Π»ΠΈΠΎΡΠ΅ΠΊΠΈ ΠΈΠ· Bacillus amyloliquefaciens ΠΈ ΠΊΠΠΠ — ΠΊΠ»ΠΎΠ½ΠΎΡΠ΅ΠΊΠΈ ΠΈΠΌΠΌΡΠ½ΠΎΠ³Π»ΠΎΠ±ΡΠ»ΠΈΠ½ΠΎΠ²ΡΡ Π³Π΅Π½ΠΎΠ² ΠΌΡΡΠΈ. ΠΠΎΠΊΠ°Π·Π°Π½ΠΎ, ΡΡΠΎ ΠΎΠΏΠΈΡΠ°Π½Π½ΡΠ΅ Π²Π΅ΠΊΡΠΎΡΠ° ΠΎΠ±Π΅ΡΠΏΠ΅ΡΠΈΠ²Π°ΡΡ Π½Π°Π΄Π΅ΠΆΠ½ΡΠΉ ΠΎΡΠ±ΠΎΡ ΠΏΠΎΠ·ΠΈΡΠΈΠ²Π½ΡΡ ΠΊΠ»ΠΎΠ½ΠΎΠ².
ΠΠ²ΡΠΎΡ Π²ΡΡΠ°ΠΆΠ°Π΅Ρ Π³Π»ΡΠ±ΠΎΠΊΡΡ Π±Π»Π°Π³ΠΎΠ΄Π°ΡΠ½ΠΎΡΡΡ ΡΠ²ΠΎΠ΅ΠΌΡ Π½Π°ΡΡΠ½ΠΎΠΌΡ ΡΡΠΊΠΎΠ²ΠΎΠ΄ΠΈΡΠ΅Π»Ρ Π΄.Π±.Π½. Π‘. Π. ΠΠ΅Π΅Π²Ρ Π·Π° Π²Π½ΠΈΠΌΠ°Π½ΠΈΠ΅, ΠΏΠΎΠ΄Π΄Π΅ΡΠΆΠΊΡ ΠΈ ΡΡΡΠΊΠΎΠ΅ ΡΡΠΊΠΎΠ²ΠΎΠ΄ΡΡΠ²ΠΎ ΡΠ°Π±ΠΎΡΠΎΠΉ.
ΠΠ²ΡΠΎΡ Π³Π»ΡΠ±ΠΎΠΊΠΎ ΠΏΡΠΈΠ·Π½Π°ΡΠ΅Π»Π΅Π½ Π·Π° ΡΠΎΡΡΡΠ΄Π½ΠΈΡΠ΅ΡΡΠ²ΠΎ Π² ΡΠ°Π±ΠΎΡΠ΅ ΠΈ ΠΏΠΎΡΡΠΎΡΠ½Π½ΡΡ Π΄ΠΎΠ±ΡΠΎΠΆΠ΅Π»Π°ΡΠ΅Π»ΡΠ½ΡΡ ΠΏΠΎΠ΄Π΄Π΅ΡΠΆΠΊΡ ΡΠ°ΡΡΠΈ ΡΠ°Π±ΠΎΡΡ, ΡΠ΄Π΅Π»Π°Π½Π½ΠΎΠΉ Π² ΠΠΈΠΎΡ ΠΈΠΌΠΈΡΠ΅ΡΠΊΠΎΠΌ ΠΠ½ΡΡΠΈΡΡΡΠ΅ Π£Π½ΠΈΠ²Π΅ΡΡΠΈΡΠ΅ΡΠ° Π³. ΠΠΈΠ»Ρ, ΠΠ°ΠΏΠ°Π΄Π½Π°Ρ ΠΠ΅ΡΠΌΠ°Π½ΠΈΡ, Π½Π΅ΠΌΠ΅ΡΠΊΠΈΠΌ ΠΊΠΎΠ»Π»Π΅Π³Π°ΠΌ: ΠΏΡΠΎΡΠ΅ΡΡΠΎΡΠ°ΠΌ: X. ΠΠ΅ΠΌΠΊΠ΅, Π. Π₯Π°Π²ΡΡΠ΅Π½Ρ, Π. ΠΠΈΠ·Π΅Π»ΡΠΌΠ°Π½Ρ, Π΄ΠΎΠΊΡΠΎΡΠ°ΠΌ: X. ΠΠ°Π½Π³Π΅, X. Π’ΠΈΡΠ·Π΅, X. Π₯Π°Π½ΡΠ΅Π½Ρ, Π. Π₯Π°ΠΌΠ°Π½Π½Ρ, Π. Π₯Π°ΠΉΠ΄ΠΎΡΠ½Ρ, Π’. ΠΠΈΡΠ΅Π»Π΅Π²ΠΎΠΉ ΠΈ Π―. ΠΠ°Π»ΡΡ , Π±ΠΈΠΎΠ»ΠΎΠ³Π°ΠΌ ΠΈ Π΄ΠΎΠΊΡΠΎΡΠ°Π½ΡΠ°ΠΌ ΠΌΠ΅Π΄ΠΈΡΠΈΠ½ΡΠΊΠΎΠ³ΠΎ ΡΠ°ΠΊΡΠ»ΡΡΠ΅ΡΠ° Π£Π½ΠΈΠ²Π΅ΡΡΠΈΡΠ΅ΡΠ° Π³. ΠΠΈΠ»Ρ: Π. ΠΠΈΡΠΎΡ, Π’. ΠΠΎΠΊΡΠΎΡΡ, Π. Π Π΅ΠΊΠ΅ ΠΈ Π‘. ΠΠΈΡΡΠΈΡ Ρ.
ΠΠ²ΡΠΎΡ Π³Π»ΡΠ±ΠΎΠΊΠΎ Π±Π»Π°Π³ΠΎΠ΄Π°ΡΠ΅Π½ Π·Π° ΡΡΠ°ΡΡΠΈΠ΅ ΠΈ Π²ΡΠ΅ΡΡΠΎΡΠΎΠ½Π½ΡΡ ΠΏΠΎΠ΄Π΄Π΅ΡΠΆΠΊΡ Π΄Π°Π½Π½ΠΎΠ³ΠΎ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΡ ΠΏΡΠΎΡΠ΅ΡΡΠΎΡΡ Π . Π₯Π°ΡΡΠ»ΠΈ (NIH, Bethesda, USA).
ΠΠ²ΡΠΎΡ Π²ΡΡΠ°ΠΆΠ°Π΅Ρ Π³Π»ΡΠ±ΠΎΠΊΡΡ Π±Π»Π°Π³ΠΎΠ΄Π°ΡΠ½ΠΎΡΡΡ ΠΏΡΠΎΡΠ΅ΡΡΠΎΡΡ, Π΄.Π±.Π½. Π. Π. ΠΠΎΠ»ΡΠ½ΠΎΠ²ΡΠΊΠΎΠΌΡ, Π΄.Π±.Π½. Π. Π. ΠΠ°Π±ΠΈΠ±ΠΎΠ²Ρ Π·Π° ΠΏΠΎΠΌΠΎΡΡ, Π΄ΠΎΠ±ΡΠΎΠΆΠ΅Π»Π°ΡΠ΅Π»ΡΠ½ΠΎΡΡΡ, ΠΏΠΎΠ»Π΅Π·Π½ΠΎΠ΅ ΠΎΠ±ΡΡΠΆΠ΄Π΅Π½ΠΈΠ΅ ΠΈ ΡΡΡΠΊΠΎΠ΅ ΠΎΡΠ½ΠΎΡΠ΅Π½ΠΈΠ΅.
ΠΠ°ΠΊΠ»ΡΡΠ΅Π½ΠΈΠ΅
.
Π’Π°ΠΊΠΈΠΌ ΠΎΠ±ΡΠ°Π·ΠΎΠΌ, ΡΠ°Π·ΡΠ°Π±ΠΎΡΠ°Π½Ρ Π΄Π²Π° Π²Π΅ΠΊΡΠΎΡΠ° Ρ ΠΏΠΎΠ·ΠΈΡΠΈΠ²Π½ΠΎΠΉ ΡΠ΅Π»Π΅ΠΊΡΠΈΠ΅ΠΉ, ΠΈΡΠΏΠΎΠ»ΡΠ·ΡΡΡΠΈΠ΅ Π»Π΅ΡΠ°Π»ΡΠ½ΡΠΉ ΡΡΡΠ΅ΠΊΡ Π³Π΅Π½Π° Π±Π°ΡΠ½Π°Π·Ρ: ΡΠΠ’440 ΠΈ ΡΠΠ° —7. ΠΠ΄ΠΈΠ½ ΠΈΠ· Π½ΠΈΡ — Π²Π΅ΠΊΡΠΎΡ ΡΠΠ’440 — ΠΏΡΠ΅Π΄Π½Π°Π·Π½Π°ΡΠ΅Π½ Π΄Π»Ρ ΡΠ΅ΡΠ΅Π½ΠΈΡ ΠΎΡΠ½ΠΎΠ²Π½ΡΡ Π·Π°Π΄Π°Ρ ΠΊΠ»ΠΎΠ½ΠΈΡΠΎΠ²Π°Π½ΠΈΡ ΠΈ ΡΡΠ±ΠΊΠ»ΠΎΠ½ΠΈΡΠΎΠ²Π°Π½ΠΈΡ ΡΡΠ°Π³ΠΌΠ΅Π½ΡΠΎΠ² ΠΠΠ, Π΄ΡΡΠ³ΠΎΠΉ — ΡΠ°Π³Π΅ΠΌΠΈΠ΄Π° ΡΠΠ° —7 — ΡΠ²Π»ΡΠ΅ΡΡΡ ΡΠΎΠ²ΡΠ΅ΠΌΠ΅Π½Π½ΡΠΌ Π²Π΅ΠΊΡΠΎΡΠΎΠΌ, ΠΏΡΠ΅Π΄ΠΎΡΡΠ°Π²Π»ΡΡΡΠΈΠΌ ΡΠΈΡΠΎΠΊΠΈΠΉ ΡΠΏΠ΅ΠΊΡΡ Π²ΠΎΠ·ΠΌΠΎΠΆΠ½ΠΎΡΡΠ΅ΠΉ Π΄Π»Ρ Π³Π΅Π½Π½ΠΎΠ³ΠΎ ΠΈΠ½ΠΆΠ΅Π½Π΅ΡΠ°.
ΠΠΎΠ·ΠΈΡΠΈΠ²Π½ΡΠΉ ΠΎΠΏΡΡ ΡΡΠΈΡ ΡΠ°Π·ΡΠ°Π±ΠΎΡΠΎΠΊ ΠΏΠΎΠ·Π²ΠΎΠ»ΡΠ΅Ρ Π·Π°ΠΊΠ»ΡΡΠΈΡΡ, ΡΡΠΎ ΡΠ°Π·ΡΠ°Π±ΠΎΡΠ°Π½Π½ΡΠΉ ΠΏΡΠΈΠ½ΡΠΈΠΏ ΡΠ΅Π»Π΅ΠΊΡΠΈΠΈ ΡΠ΅ΠΊΠΎΠΌΠ±ΠΈΠ½Π°Π½ΡΠ½ΡΡ ΠΊΠ»ΠΎΠ½ΠΎΠ² Ρ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠ΅ΠΌ ΡΡΠ»ΠΎΠ²Π½ΠΎΠ»Π΅ΡΠ°Π»ΡΠ½ΠΎΠ³ΠΎ Π³Π΅Π½Π° Π±Π°ΡΠ½Π°Π·Ρ ΠΌΠΎΠΆΠ΅Ρ Π»Π΅ΡΡ Π² ΠΎΡΠ½ΠΎΠ²Ρ ΡΠΎΠ·Π΄Π°Π½ΠΈΡ Π½ΠΎΠ²ΠΎΠ³ΠΎ ΠΏΠΎΠΊΠΎΠ»Π΅Π½ΠΈΡ Π²Π΅ΠΊΡΠΎΡΠΎΠ² Ρ Π½ΡΠ»Π΅Π²ΡΠΌ ΡΠΎΠ½ΠΎΠΌ.
Π‘ΠΏΠΈΡΠΎΠΊ Π»ΠΈΡΠ΅ΡΠ°ΡΡΡΡ
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- Yazynin SA, Deyev SM, Jucovic M and Hartley RW, A plasmid vector with positive selection and directional cloning based on a conditionally lethal gene. Gene 159(1): 131−2, 1996.
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