Π Π΅ΠΏΠ»ΠΈΠΊΠ°ΡΠΈΠΎΠ½Π½ΡΠΉ Π±Π΅Π»ΠΎΠΊ RC ΠΏΠ»Π°Π·ΠΌΠΈΠ΄Ρ pNB2 ΡΠ΅ΡΠΌΠΎΡΠΈΠ»ΡΠ½ΡΡ Π±Π°ΠΊΡΠ΅ΡΠΈΠΉ Clostridium thermosaccharolyticum: ΠΠ΄Π΅Π½ΡΠΈΡΠΈΠΊΠ°ΡΠΈΡ, ΡΠΊΡΠΏΡΠ΅ΡΡΠΈΡ ΠΈ ΠΌΡΡΠ°ΡΠΈΠΎΠ½Π½ΡΠΉ Π°Π½Π°Π»ΠΈΠ·
![ΠΠΈΡΡΠ΅ΡΡΠ°ΡΠΈΡ: Π Π΅ΠΏΠ»ΠΈΠΊΠ°ΡΠΈΠΎΠ½Π½ΡΠΉ Π±Π΅Π»ΠΎΠΊ RC ΠΏΠ»Π°Π·ΠΌΠΈΠ΄Ρ pNB2 ΡΠ΅ΡΠΌΠΎΡΠΈΠ»ΡΠ½ΡΡ
Π±Π°ΠΊΡΠ΅ΡΠΈΠΉ Clostridium thermosaccharolyticum: ΠΠ΄Π΅Π½ΡΠΈΡΠΈΠΊΠ°ΡΠΈΡ, ΡΠΊΡΠΏΡΠ΅ΡΡΠΈΡ ΠΈ ΠΌΡΡΠ°ΡΠΈΠΎΠ½Π½ΡΠΉ Π°Π½Π°Π»ΠΈΠ·](https://niscu.ru/work/3748939/cover.png)
Π ΠΏΠΎΡΠ»Π΅Π΄Π½ΠΈΠ΅ Π³ΠΎΠ΄Ρ Π΄ΠΎΡΡΠΈΠ³Π½ΡΡΡ Π²ΠΏΠ΅ΡΠ°ΡΠ»ΡΡΡΠΈΠ΅ ΡΡΠΏΠ΅Ρ ΠΈ ΠΏΠΎ ΠΊΠ»ΠΎΠ½ΠΈΡΠΎΠ²Π°Π½ΠΈΡ Π±Π°ΠΊΡΠ΅ΡΠΈΠ°Π»ΡΠ½ΡΡ Π³Π΅Π½ΠΎΠ² Π²ΠΎ Π²ΡΠ΅ΡΡΠΎΡΠΎΠ½Π½Π΅ ΠΈΠ·ΡΡΠ΅Π½Π½ΡΡ Π±Π°ΠΊΡΠ΅ΡΠΈΡΡ Escherichia coli ΠΈ Bacillus subtilis ΠΈ Π°Π΄Π°ΠΏΡΠΈΡΠΎΠ²Π°Π½Π½ΡΡ Π΄Π»Ρ Π½ΠΈΡ Π²Π΅ΠΊΡΠΎΡΠ°Ρ . ΠΠ΅ΡΠΌΠΎΡΡΡ Π½Π° ΡΡΠΎ ΠΎΡΡΠ°Π΅ΡΡΡ Π²Π΅ΡΡΠΌΠ° Π°ΠΊΡΡΠ°Π»ΡΠ½ΠΎΠΉ ΠΏΡΠΎΠ±Π»Π΅ΠΌΠ° ΠΏΠΎΠΈΡΠΊΠ° ΠΈ ΠΈΠ·ΡΡΠ΅Π½ΠΈΡ Π½ΠΎΠ²ΡΡ ΠΏΠ»Π°Π·ΠΌΠΈΠ΄, ΠΊΠΎΡΠΎΡΡΠ΅ ΠΌΠΎΠ³ΡΡ Π±ΡΡΡ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½Ρ Π² ΠΊΠ°ΡΠ΅ΡΡΠ²Π΅ Π³Π΅Π½Π½ΠΎ-ΠΈΠ½ΠΆΠ΅Π½Π΅ΡΠ½ΡΡ Π²Π΅ΠΊΡΠΎΡΠΎΠ² Π² ΡΠΏΠ΅ΡΠΈΠ°Π»ΠΈΠ·ΠΈΡΠΎΠ²Π°Π½Π½ΡΡ Π³ΡΡΠΏΠΏΠ°Ρ Π±Π°ΠΊΡΠ΅ΡΠΈΠΉ. Π ΡΠ²ΡΠ·ΠΈ Ρ ΡΡΠΈΠΌ… 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 ΡΠΎΠΊΡΠ°ΡΠ΅Π½ΠΈΠΉ
- I. ΠΠΠ’ΠΠ ΠΠ’Π£Π ΠΠ«Π ΠΠΠΠΠ
- 1. 1. ΠΠ΅Ρ Π°Π½ΠΈΠ·ΠΌ ΡΠ΅ΠΏΠ»ΠΈΠΊΠ°ΡΠΈΠΈ rolling circle
- 1. 2. ΠΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠ΅ ΡΠΈΡΡΠ΅ΠΌΡ «ΡΡΠ°Π±ΠΈΠ»ΡΠ½ΡΠΉ ΡΠΎΠΊΡΠΈΠ½-Π»Π°Π±ΠΈΠ»ΡΠ½ΡΠΉ Π°Π½ΡΠΈΡΠΎΠΊΡΠΈΠ½» Π΄Π»Ρ ΠΏΠΎΠ΄Π΄Π΅ΡΠΆΠ°Π½ΠΈΡ ΠΏΠ»Π°Π·ΠΌΠΈΠ΄ Π² ΠΊΠ»Π΅ΡΠΊΠ°Ρ Π±Π°ΠΊΡΠ΅ΡΠΈΠΉ
- 1. 3. ΠΠΊΡΠΏΡΠ΅ΡΡΠΈΡ ΡΠ΅ΠΊΠΎΠΌΠ±ΠΈΠ½Π°Π½ΡΠ½ΡΡ Π³Π΅Π½ΠΎΠ² Π² Π. coli. ΠΡΠΎΠ±Π΅Π½Π½ΠΎΡΡΠΈ ΡΠ°Π±ΠΎΡΡ Ρ ΡΠΎΠΊΡΠΈΡΠ½ΡΠΌΠΈ Π³Π΅Π½Π°ΠΌΠΈ
- ΠΠΠ‘ΠΠΠ ΠΠΠΠΠ’ΠΠΠ¬ΠΠΠ― Π§ΠΠ‘Π’
- II. ΠΠΠ’ΠΠ ΠΠΠΠ« Π ΠΠΠ’ΠΠΠ«
- III. ΠΠΠ‘Π£ΠΠΠΠΠΠ Π ΠΠΠ£ΠΠ¬Π’ΠΠ’ΠΠ
- III. 1 ΠΠ½Π°Π»ΠΈΠ· Π½ΡΠΊΠ»Π΅ΠΎΡΠΈΠ΄Π½ΠΎΠΉ ΠΏΠΎΡΠ»Π΅Π΄ΠΎΠ²Π°ΡΠ΅Π»ΡΠ½ΠΎΡΡΠΈ pNB
- 111. 2. ΠΠ·ΠΌΠ΅ΡΠ΅Π½ΠΈΠ΅ ΠΏΡΠΎΠΌΠΎΡΠΎΡΠ½ΠΎΠΉ Π°ΠΊΡΠΈΠ²Π½ΠΎΡΡΠΈ ΡΡΠ°Π³ΠΌΠ΅Π½ΡΠΎΠ² ΠΏΠ»Π°Π·ΠΌΠΈΠ΄Ρ pNB
- 111. 3. ΠΠ½Π°Π»ΠΈΠ· Π°ΠΌΠΈΠ½ΠΎΠΊΠΈΡΠ»ΠΎΡΠ½ΠΎΠΉ ΠΏΠΎΡΠ»Π΅Π΄ΠΎΠ²Π°ΡΠ΅Π»ΡΠ½ΠΎΡΡΠΈ Π±Π΅Π»ΠΊΠ°, ΡΠΎΠΎΡΠ²Π΅ΡΡΡΠ²ΡΡΡΠ΅Π³ΠΎ ΠΠ Π‘289.'
- 111. 4. Π Π΅Π³ΠΈΡΡΡΠ°ΡΠΈΡ ΠΎΠ΄Π½ΠΎΠ½ΠΈΡΠ΅Π²ΠΎΠΉ ΡΠΎΡΠΌΡ ΠΏΠ»Π°Π·ΠΌΠΈΠ΄Ρ pNB2 Π² ΠΊΠ»Π΅ΡΠΊΠ°Ρ
- Clostridium thermosaccharolyticum
- 111. 5. ΠΠΊΡΠΏΡΠ΅ΡΡΠΈΡ repN Π³Π΅Π½Π°
- III. 5.1 ΠΠΎΠ΄ΠΈΡΠΈΠΊΠ°ΡΠΈΡ Π½Π΅ ΡΡΡΠ΅ΠΊΡΠΈΠ²Π½ΡΡ «Π½Π΅-ATG» ΠΈΠ½ΠΈΡΠΈΠΈΡΡΡΡΠΈΡ ΠΊΠΎΠ΄ΠΎΠ½ΠΎΠ² repN Π³Π΅Π½Π°
- III. 5.2. ΠΠΊΡΠΏΡΠ΅ΡΡΠΈΡ repN tqwl Π² Π±Π΅ΡΠΊΠ»Π΅ΡΠΎΡΠ½ΠΎΠΉ ΡΠΈΡΡΠ΅ΠΌΠ΅ ΡΡΠ°Π½ΡΠ»ΡΡΠΈΠΈ ΡΡΠ°Π½ΡΠΊΡΠΈΠΏΡΠΈΠΈ S
- 111. 5. 3. Π‘ΠΎΠ·Π΄Π°Π½ΠΈΠ΅ ΡΠ΅ΠΊΠΎΠΌΠ±ΠΈΠ½Π°Π½ΡΠ½ΠΎΠΉ ΠΏΠ»Π°Π·ΠΌΠΈΠ΄Ρ Π΄Π»Ρ ΡΠΎΡΠΊΡΠΏΡΠ΅ΡΡΠΈΠΈ ΡΠ΅Π΄ΠΊΠΎΠΉ Π² Π. coli ΡΠ ΠΠ
- 111. 5. 4. Π’ΡΠ°Π½ΡΡΠΎΡΠΌΠ°ΡΠΈΡ ΠΊΠ»Π΅ΡΠΎΠΊ E. coli ΡΠ΅ΠΊΠΎΠΌΠ±ΠΈΠ½Π°Π½ΡΠ½ΡΠΌΠΈ ΠΏΠ»Π°Π·ΠΌΠΈΠ΄Π°ΠΌΠΈ, ΡΠΎΠ΄Π΅ΡΠΆΠ°ΡΠΈΠΌΠΈ repN Π³Π΅Π½
- 111. 5. 5. ΠΠΊΡΠΏΡΠ΅ΡΡΠΈΡ repN Π³Π΅Π½Π° in vivo Π² E. col
- III. 6. ΠΡΠΈΡΡΠΊΠ° RepN Π±Π΅Π»ΠΊΠ° ΠΈΠ· Π»ΠΈΠ·Π°ΡΠΎΠ² E. col
- III. 6.1 ΠΠΎΠ΄ΠΈΡΠΈΠΊΠ°ΡΠΈΡ 5' ΠΊΠΎΠ½ΡΠ° repN Π³Π΅Π½Π° ΠΏΠΎΡΠ»Π΅Π΄ΠΎΠ²Π°ΡΠ΅Π»ΡΠ½ΠΎΡΡΡΡ, ΡΠΎΠΎΡΠ²Π΅ΡΡΡΠ²ΡΡΡΠ΅ΠΉ ΡΠ΅ΡΡΠΈ Π³ΠΈΡΡΠΈΠ΄ΠΈΠ½Π°ΠΌ
- 111. 6. 2. ΠΠΊΡΠΏΡΠ΅ΡΡΠΈΡ Π³Π΅ΡΡΠ£Π³Π΅Π½Π°, ΠΌΠΎΠ΄ΠΈΡΠΈΡΠΈΡΠΎΠ²Π°Π½Π½ΠΎΠ³ΠΎ ΠΏΠΎΡΠ»Π΅Π΄ΠΎΠ²Π°ΡΠ΅Π»ΡΠ½ΠΎΡΡΡΡ ΡΠΎΠΎΡΠ²Π΅ΡΡΡΠ²ΡΡΡΠ΅ΠΉ ΡΠ΅ΡΡΠΈ Π³ΠΈΡΡΠΈΠ΄ΠΈΠ½Π°ΠΌ
- 111. 6. 3. ΠΡΠΈΡΡΠΊΠ° His-repN Π±Π΅Π»ΠΊΠ° ΠΈΠ· Π½Π΅ΡΠ°ΡΡΠ²ΠΎΡΠΈΠΌΠΎΠΉ ΡΡΠ°ΠΊΡΠΈΠΈ
- 111. 6. 4. ΠΠΎΠ΄Π±ΠΎΡ ΡΡΠ»ΠΎΠ²ΠΈΠΉ Π΄Π»Ρ ΡΠ²Π΅Π»ΠΈΡΠ΅Π½ΠΈΡ ΡΠ°ΡΡΠ²ΠΎΡΠΈΠΌΠΎΡΡΠΈ Π±Π΅Π»ΠΊΠ° in vivo
- III. 7. ΠΠ·ΡΡΠ΅Π½ΠΈΠ΅ ΠΏΡΠΈΡΠΈΠ½ Π»Π΅ΡΠ°Π»ΡΠ½ΠΎΠ³ΠΎ Π΄Π΅ΠΉΡΡΠ²ΠΈΡ RepN Π±Π΅Π»ΠΊΠ° Π½Π° ΠΊΠ»Π΅ΡΠΊΠΈ Π. col
- 111. 7. 1. Π‘Π°ΠΉΡ-Π½Π°ΠΏΡΠ°Π²Π»Π΅Π½Π½ΡΠΉ ΠΌΡΡΠ°Π³Π΅Π½Π΅Π· Π°ΠΌΠΈΠ½ΠΎΠΊΠΈΡΠ»ΠΎΡ, Ρ Π°ΡΠ°ΠΊΡΠ΅ΡΠ½ΡΡ Π΄Π»Ρ Π°ΠΊΡΠΈΠ²Π½ΠΎΠ³ΠΎ ΡΠ΅Π½ΡΡΠ° ΡΠ΅Π»Π°ΠΊΡΠ°Π·
- 111. 7. 2. ΠΠ΅Π»Π΅ΡΠΈΠΎΠ½Π½ΠΎΠ΅ ΠΊΠ°ΡΡΠΈΡΠΎΠ²Π°Π½ΠΈΠ΅ ΠΎΠ±Π»Π°ΡΡΠΈ RepN Π±Π΅Π»ΠΊΠ°, ΠΎΡΠ²Π΅ΡΡΡΠ²Π΅Π½Π½ΠΎΠΉ Π·Π° Π΅Π³ΠΎ Π»Π΅ΡΠ°Π»ΡΠ½ΠΎΠ΅ Π΄Π΅ΠΉΡΡΠ²ΠΈΠ΅
- ΠΠ«ΠΠΠΠ«
Π Π΅ΠΏΠ»ΠΈΠΊΠ°ΡΠΈΠΎΠ½Π½ΡΠΉ Π±Π΅Π»ΠΎΠΊ RC ΠΏΠ»Π°Π·ΠΌΠΈΠ΄Ρ pNB2 ΡΠ΅ΡΠΌΠΎΡΠΈΠ»ΡΠ½ΡΡ Π±Π°ΠΊΡΠ΅ΡΠΈΠΉ Clostridium thermosaccharolyticum: ΠΠ΄Π΅Π½ΡΠΈΡΠΈΠΊΠ°ΡΠΈΡ, ΡΠΊΡΠΏΡΠ΅ΡΡΠΈΡ ΠΈ ΠΌΡΡΠ°ΡΠΈΠΎΠ½Π½ΡΠΉ Π°Π½Π°Π»ΠΈΠ· (ΡΠ΅ΡΠ΅ΡΠ°Ρ, ΠΊΡΡΡΠΎΠ²Π°Ρ, Π΄ΠΈΠΏΠ»ΠΎΠΌ, ΠΊΠΎΠ½ΡΡΠΎΠ»ΡΠ½Π°Ρ)
ΠΠ°ΠΊΡΠ΅ΡΠΈΠΈ Clostridium thermosaccharolyticum ΡΠ²Π»ΡΡΡΡΡ ΡΠ΅ΡΠΌΠΎΡΠΈΠ»ΡΠ½ΡΠΌΠΈ Π°Π½Π°ΡΡΠΎΠ±Π½ΡΠΌΠΈ ΠΌΠΈΠΊΡΠΎΠΎΡΠ³Π°Π½ΠΈΠ·ΠΌΠ°ΠΌΠΈ. ΠΠ½ΠΎΠ³ΠΈΠ΅ Π²ΠΈΠ΄Ρ ΡΠΎΠ΄Π° Clostridium ΠΏΡΠ΅Π΄ΡΡΠ°Π²Π»ΡΡΡ Π±ΠΎΠ»ΡΡΠΎΠΉ ΠΈΠ½ΡΠ΅ΡΠ΅Ρ Π΄Π»Ρ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΡ Π² Π±ΠΈΠΎΡΠ΅Ρ Π½ΠΎΠ»ΠΎΠ³ΠΈΡΠ΅ΡΠΊΠΈΡ ΡΠ΅Π»ΡΡ . Π‘ΡΠ΅Π΄ΠΈ ΠΏΡΠΎΠ΄ΡΠΊΡΠΎΠ² ΠΈΡ ΠΌΠ΅ΡΠ°Π±ΠΎΠ»ΠΈΠ·ΠΌΠ° ΠΈΠΌΠ΅ΡΡΡΡ ΠΏΠ΅ΡΡΠΏΠ΅ΠΊΡΠΈΠ²Π½ΡΠ΅ ΡΠ½Π΅ΡΠ³ΠΎΠ½ΠΎΡΠΈΡΠ΅Π»ΠΈ (ΡΡΠ°Π½ΠΎΠ», Π±ΡΡΠ°Π½ΠΎΠ», Π²ΠΎΠ΄ΠΎΡΠΎΠ΄), ΠΎΡΠ³Π°Π½ΠΈΡΠ΅ΡΠΊΠΈΠ΅ ΠΊΠΈΡΠ»ΠΎΡΡ (ΡΠΊΡΡΡΠ½Π°Ρ, ΠΏΡΠΎΠΏΠΈΠΎΠ½ΠΎΠ²Π°Ρ, ΠΌΠ°ΡΠ»ΡΠ½Π°Ρ), ΡΠ΅Ρ Π½ΠΎΠ»ΠΎΠ³ΠΈΡΠ΅ΡΠΊΠΈ Π²Π°ΠΆΠ½ΡΠ΅ ΡΠ΅ΡΠΌΠ΅Π½ΡΡ (ΡΠ΅Π»Π»ΡΠ»Π°Π·Ρ, Π³Π΅ΠΌΠΈΡΠ΅Π»Π»ΡΠ»Π°Π·Ρ, ΠΈ Ρ. Π΄.). ΠΠΎ ΠΎΡΠ½ΠΎΡΠΈΡΠ΅Π»ΡΠ½ΠΎ Π½ΠΈΠ·ΠΊΠΈΠ΅ ΡΠΊΠΎΡΠΎΡΡΠΈ ΡΠΎΡΡΠ° ΠΊΠ»ΠΎΡΡΡΠΈΠ΄ΠΈΠΉ, Π½Π΅ΠΎΠ±Ρ ΠΎΠ΄ΠΈΠΌΠΎΡΡΡ ΡΡΡΠΎΠ³ΠΎΠ³ΠΎ ΡΠΎΠ±Π»ΡΠ΄Π΅Π½ΠΈΡ Π°Π½Π°ΡΡΠΎΠ±Π½ΡΡ ΡΡΠ»ΠΎΠ²ΠΈΠΉ, Π½Π΅Π²ΡΡΠΎΠΊΠΈΠ΅ Π²ΡΡ ΠΎΠ΄Ρ ΡΠ΅Π»Π΅Π²ΡΡ ΠΏΡΠΎΠ΄ΡΠΊΡΠΎΠ² ΡΠ΄Π΅ΡΠΆΠΈΠ²Π°ΡΡ ΠΏΡΠ°ΠΊΡΠΈΡΠ΅ΡΠΊΠΎΠ΅ ΠΏΡΠΈΠΌΠ΅Π½Π΅Π½ΠΈΠ΅ ΡΡΠΈΡ Π±Π°ΠΊΡΠ΅ΡΠΈΠΉ. Π‘ Π΄ΡΡΠ³ΠΎΠΉ ΡΡΠΎΡΠΎΠ½Ρ, Π½Π΅ΠΊΠΎΡΠΎΡΡΠ΅ ΠΏΡΠ΅Π΄ΡΡΠ°Π²ΠΈΡΠ΅Π»ΠΈ ΡΠΎΠ΄Π° Clostridium ΠΈΠΌΠ΅ΡΡ Π±ΠΎΠ»ΡΡΠΎΠ΅ ΠΌΠ΅Π΄ΠΈΡΠΈΠ½ΡΠΊΠΎΠ΅ Π·Π½Π°ΡΠ΅Π½ΠΈΠ΅, ΡΠ°ΠΊ ΠΊΠ°ΠΊ ΡΠ²Π»ΡΡΡΡΡ ΠΏΡΠΎΠ΄ΡΡΠ΅Π½ΡΠ°ΠΌΠΈ ΡΠΈΠ»ΡΠ½ΠΎΠ΄Π΅ΠΉΡΡΠ²ΡΡΡΠΈΡ ΡΠΎΠΊΡΠΈΠ½ΠΎΠ² (ΡΡΠΎΠ»Π±Π½ΡΡΠ½ΡΠΉ ΠΈ Π±ΠΎΡΡΠ»ΠΈΠ½ΠΈΡΠ΅ΡΠΊΠΈΠΉ ΡΠΎΠΊΡΠΈΠ½Ρ).
Π ΠΏΠΎΡΠ»Π΅Π΄Π½ΠΈΠ΅ Π³ΠΎΠ΄Ρ Π΄ΠΎΡΡΠΈΠ³Π½ΡΡΡ Π²ΠΏΠ΅ΡΠ°ΡΠ»ΡΡΡΠΈΠ΅ ΡΡΠΏΠ΅Ρ ΠΈ ΠΏΠΎ ΠΊΠ»ΠΎΠ½ΠΈΡΠΎΠ²Π°Π½ΠΈΡ Π±Π°ΠΊΡΠ΅ΡΠΈΠ°Π»ΡΠ½ΡΡ Π³Π΅Π½ΠΎΠ² Π²ΠΎ Π²ΡΠ΅ΡΡΠΎΡΠΎΠ½Π½Π΅ ΠΈΠ·ΡΡΠ΅Π½Π½ΡΡ Π±Π°ΠΊΡΠ΅ΡΠΈΡΡ Escherichia coli ΠΈ Bacillus subtilis ΠΈ Π°Π΄Π°ΠΏΡΠΈΡΠΎΠ²Π°Π½Π½ΡΡ Π΄Π»Ρ Π½ΠΈΡ Π²Π΅ΠΊΡΠΎΡΠ°Ρ . ΠΠ΅ΡΠΌΠΎΡΡΡ Π½Π° ΡΡΠΎ ΠΎΡΡΠ°Π΅ΡΡΡ Π²Π΅ΡΡΠΌΠ° Π°ΠΊΡΡΠ°Π»ΡΠ½ΠΎΠΉ ΠΏΡΠΎΠ±Π»Π΅ΠΌΠ° ΠΏΠΎΠΈΡΠΊΠ° ΠΈ ΠΈΠ·ΡΡΠ΅Π½ΠΈΡ Π½ΠΎΠ²ΡΡ ΠΏΠ»Π°Π·ΠΌΠΈΠ΄, ΠΊΠΎΡΠΎΡΡΠ΅ ΠΌΠΎΠ³ΡΡ Π±ΡΡΡ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½Ρ Π² ΠΊΠ°ΡΠ΅ΡΡΠ²Π΅ Π³Π΅Π½Π½ΠΎ-ΠΈΠ½ΠΆΠ΅Π½Π΅ΡΠ½ΡΡ Π²Π΅ΠΊΡΠΎΡΠΎΠ² Π² ΡΠΏΠ΅ΡΠΈΠ°Π»ΠΈΠ·ΠΈΡΠΎΠ²Π°Π½Π½ΡΡ Π³ΡΡΠΏΠΏΠ°Ρ Π±Π°ΠΊΡΠ΅ΡΠΈΠΉ. Π ΡΠ²ΡΠ·ΠΈ Ρ ΡΡΠΈΠΌ Π±ΠΎΠ»ΡΡΠΎΠΉ ΠΈΠ½ΡΠ΅ΡΠ΅Ρ ΠΏΡΠ΅Π΄ΡΡΠ°Π²Π»ΡΠ΅Ρ ΡΠ°ΠΊΠΆΠ΅ ΠΈΠ·ΡΡΠ΅Π½ΠΈΠ΅ Π±Π΅Π»ΠΊΠΎΠ², ΡΡΠ°ΡΡΠ²ΡΡΡΠΈΡ Π² ΡΠ΅ΠΏΠ»ΠΈΠΊΠ°ΡΠΈΠΈ ΠΈΡΡΠ»Π΅Π΄ΡΠ΅ΠΌΡΡ ΠΏΠ»Π°Π·ΠΌΠΈΠ΄. ΠΡΠΎΠ±Π΅Π½Π½ΠΎ ΡΡΡΠ΅ΡΡΠ²Π΅Π½Π½ΡΠΌ ΡΠ²Π»ΡΠ΅ΡΡΡ ΠΏΠΎΠ΄Π±ΠΎΡ Π½ΠΎΠ²ΡΡ ΡΠΈΡΡΠ΅ΠΌ Π΄Π»Ρ ΠΊΠ»ΠΎΠ½ΠΈΡΠΎΠ²Π°Π½ΠΈΡ Π³Π΅Π½ΠΎΠ² ΠΈΠ· ΡΠ΅ΡΠΌΠΎΡΠΈΠ»ΡΠ½ΡΡ Π±Π°ΠΊΡΠ΅ΡΠΈΠΉ, ΡΠ°ΠΊ ΠΊΠ°ΠΊ ΠΌΠ½ΠΎΠ³ΠΈΠ΅ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°ΡΠ΅Π»ΠΈ ΡΡΠ°Π»ΠΊΠΈΠ²Π°ΡΡΡΡ Ρ ΡΡΡΠ΄Π½ΠΎΡΡΡΠΌΠΈ ΠΏΡΠ°Π²ΠΈΠ»ΡΠ½ΠΎΠ³ΠΎ ΡΠΎΠ»Π΄ΠΈΠ½Π³Π° ΡΠ΅ΠΊΠΎΠΌΠ±ΠΈΠ½Π°Π½ΡΠ½ΡΡ Π±Π΅Π»ΠΊΠΎΠ² ΡΠ΅ΡΠΌΠΎΡΠΈΠ»ΡΠ½ΡΡ ΠΎΡΠ³Π°Π½ΠΈΠ·ΠΌΠΎΠ² Π² ΡΠΈΡΠΎΠΊΠΎ ΠΈΡΠΏΠΎΠ»ΡΠ·ΡΠ΅ΠΌΡΡ Π±Π°ΠΊΡΠ΅ΡΠΈΡΡ E. coli ΠΈ Π.subtilis.
Π£Π΄ΠΈΠ²ΠΈΡΠ΅Π»ΡΠ½ΠΎΠΉ ΠΎΡΠΎΠ±Π΅Π½Π½ΠΎΡΡΡΡ ΡΠ΅ΡΠΌΠΎΡΠΈΠ»ΡΠ½ΡΡ ΠΊΠ»ΠΎΡΡΡΠΈΠ΄ΠΈΠΉ ΡΠ²Π»ΡΠ΅ΡΡΡ Π½ΠΈΠ·ΠΊΠΎΠ΅ ΡΠΎΠ΄Π΅ΡΠΆΠ°Π½ΠΈΠ΅ «ΡΡΠ³ΠΎΠΏΠ»Π°Π²ΠΊΠΈΡ » GC-nap (ΠΎΠΊΠΎΠ»ΠΎ 38%), ΡΡΠΎ Π½Π΅ ΠΊΠΎΡΡΠ΅Π»ΠΈΡΡΠ΅Ρ Ρ Π²ΡΡΠΎΠΊΠΎΠΉ ΡΠΎΠ»Π΅ΡΠ°Π½ΡΠ½ΠΎΡΡΡΡ ΡΡΠΈΡ Π±Π°ΠΊΡΠ΅ΡΠΈΠΉ ΠΊ ΡΠ΅ΠΌΠΏΠ΅ΡΠ°ΡΡΡΠ΅ (ΠΎΠΏΡΠΈΠΌΡΠΌ ΡΠΎΡΡΠ° 60−65Β°Π‘) ΠΈ ΠΎΡΠ»ΠΈΡΠ°Π΅Ρ ΠΈΡ ΠΎΡ Π΄ΡΡΠ³ΠΈΡ ΡΠ΅ΡΠΌΠΎΡΠΈΠ»ΡΠ½ΡΡ ΠΎΡΠ³Π°Π½ΠΈΠ·ΠΌΠΎΠ² [1]. ΠΠ»Π°ΡΡΠΈΡΠ΅ΡΠΊΠ°Ρ Π·Π°ΠΊΠΎΠ½ΠΎΠΌΠ΅ΡΠ½ΠΎΡΡΡ ΠΏΡΠ΅Π΄ΠΏΠΎΠ»Π°Π³Π°Π΅Ρ ΡΠ²Π΅Π»ΠΈΡΠ΅Π½ΠΈΠ΅ GC-ΡΠΎΡΡΠ°Π²Π° ΠΠΠ Ρ ΠΏΠΎΠ²ΡΡΠ΅Π½ΠΈΠ΅ΠΌ ΡΠ΅ΠΌΠΏΠ΅ΡΠ°ΡΡΡΠ½ΠΎΠ³ΠΎ ΠΎΠΏΡΠΈΠΌΡΠΌΠ° ΠΆΠΈΠ·Π½Π΅Π΄Π΅ΡΡΠ΅Π»ΡΠ½ΠΎΡΡΠΈ Π±Π°ΠΊΡΠ΅ΡΠΈΠΉ. Π’Π°ΠΊ Π½Π΅ΠΊΠΎΡΠΎΡΡΠ΅ ΠΏΡΠ΅Π΄ΡΡΠ°Π²ΠΈΡΠ΅Π»ΠΈ ΡΠ΅ΡΠΌΠΎΡΠΈΠ»ΡΠ½ΡΡ Π±Π°ΠΊΡΠ΅ΡΠΈΠΉ ΡΠΎΠ΄Π° Thermus ΠΈΠΌΠ΅ΡΡ GC-ΡΠΎΡΡΠ°Π² ΡΠ²ΡΡΠ΅ 70% [2], Π² ΡΠΎ Π²ΡΠ΅ΠΌΡ ΠΊΠ°ΠΊ Π² ΠΊΠ»Π΅ΡΠΊΠ°Ρ ΡΠΈΠΏΠΈΡΠ½ΠΎ ΠΌΠ΅Π·ΠΎΡΠΈΠ»ΡΠ½ΡΡ E. coli ΠΎΠ½ ΡΠΎΡΡΠ°Π²Π»ΡΠ΅Ρ 51% [3]. Π ΡΠ²ΡΠ·ΠΈ Ρ ΡΡΠΈΠΌ ΠΏΡΠ΅Π΄ΡΡΠ°Π²Π»ΡΠ΅Ρ ΠΈΠ½ΡΠ΅ΡΠ΅Ρ ΠΈΠ·ΡΡΠ΅Π½ΠΈΠ΅ ΠΎΡΠΎΠ±Π΅Π½Π½ΠΎΡΡΠ΅ΠΉ ΡΠ΅ΠΏΠ»ΠΈΠΊΠ°ΡΠΈΠΈ ΠΠΠ Π² ΡΠ΅ΡΠΌΠΎΡΠΈΠ»ΡΠ½ΡΡ ΠΊΠ»ΠΎΡΡΡΠΈΠ΄ΠΈΡΡ .
Π ΡΠ°Π±ΠΎΡΠ΅ ΠΠ΅Π»ΠΎΠ³ΡΡΠΎΠ²ΠΎΠΉ Π. Π. Ρ ΡΠΎΠ°Π²ΡΠΎΡΠ°ΠΌΠΈ [4] Π² ΠΊΠ»Π΅ΡΠΊΠ°Ρ Π‘. thermosaccarolyticum DSM571 Π±ΡΠ»ΠΈ ΠΈΠ΄Π΅Π½ΡΠΈΡΠΈΡΠΈΡΠΎΠ²Π°Π½Ρ Π΄Π²Π΅ Π½Π΅Π±ΠΎΠ»ΡΡΠΈΠ΅ ΠΏΠ»Π°Π·ΠΌΠΈΠ΄Ρ pNBl ΠΈ pNB2 Ρ Π½Π΅ΠΈΠ·Π²Π΅ΡΡΠ½ΡΠΌΠΈ ΡΡΠ½ΠΊΡΠΈΡΠΌΠΈ. ΠΠ½Π°Π»ΠΈΠ· ΠΏΠΎΠ»Π½ΠΎΠΉ Π½ΡΠΊΠ»Π΅ΠΎΡΠΈΠ΄Π½ΠΎΠΉ ΠΏΠΎΡΠ»Π΅Π΄ΠΎΠ²Π°ΡΠ΅Π»ΡΠ½ΠΎΡΡΠΈ pNB2 ΠΏΠΎΠΊΠ°Π·Π°Π», ΡΡΠΎ ΡΡΠ° ΠΏΠ»Π°Π·ΠΌΠΈΠ΄Π° ΡΠΎΡΡΠΎΠΈΡ ΠΈΠ· 1882 ΠΏΠ°Ρ Π½ΡΠΊΠ»Π΅ΠΎΡΠΈΠ΄ΠΎΠ² ΠΈ ΡΠ²Π»ΡΠ΅ΡΡΡ Π΄Π΅Π»Π΅ΡΠΈΡΠΎΠ²Π°Π½Π½ΠΎΠΉ ΡΠΎΡΠΌΠΎΠΉ ΠΏΠ»Π°Π·ΠΌΠΈΠ΄Ρ pNBl, ΠΏΡΠΈΡΠ΅ΠΌ ΡΠΎΠ΄Π΅ΡΠΆΠ°Π½ΠΈΠ΅ Π² Π½Π΅ΠΉ GC ΠΏΠ°Ρ ΡΠΎΡΡΠ°Π²Π»ΡΠ΅Ρ Π²ΡΠ΅Π³ΠΎ 27,2%. Π‘ΡΠΎΠ»Ρ Π½ΠΈΠ·ΠΊΠΎΠ΅ ΡΠΎΠ΄Π΅ΡΠΆΠ°Π½ΠΈΠ΅ «ΡΡΠ³ΠΎΠΏΠ»Π°Π²ΠΊΠΈΡ » GC ΠΏΠ°Ρ Π΄Π°Π΅Ρ Π²ΠΎΠ·ΠΌΠΎΠΆΠ½ΠΎΡΡΡ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°ΡΡ ΡΡΡ ΠΏΠ»Π°Π·ΠΌΠΈΠ΄Ρ ΠΊΠ°ΠΊ ΡΠ΄ΠΎΠ±Π½ΡΡ ΠΌΠΎΠ΄Π΅Π»Ρ Π΄Π»Ρ ΠΈΠ·ΡΡΠ΅Π½ΠΈΡ ΠΌΠ΅Ρ Π°Π½ΠΈΠ·ΠΌΠΎΠ² ΡΠ΅ΠΏΠ»ΠΈΠΊΠ°ΡΠΈΠΈ ΠΈ ΡΡΠ°Π±ΠΈΠ»ΠΈΠ·Π°ΡΠΈΠΈ ΠΠ’-Π±ΠΎΠ³Π°ΡΠΎΠΉ ΠΠΠ Π² Π±Π°ΠΊΡΠ΅ΡΠΈΠ°Π»ΡΠ½ΠΎΠΉ ΠΊΠ»Π΅ΡΠΊΠ΅.
ΠΠ°ΡΡΠΎΡΡΠ°Ρ ΡΠ°Π±ΠΎΡΠ° Π±ΡΠ»Π° ΠΏΡΠ΅Π΄ΠΏΡΠΈΠ½ΡΡΠ° Ρ ΡΠ΅Π»ΡΡ Π²ΡΠ΅ΡΡΠΎΡΠΎΠ½Π½Π΅Π³ΠΎ ΠΈΠ·ΡΡΠ΅Π½ΠΈΡ ΠΊΠ»ΡΡΠ΅Π²ΠΎΠ³ΠΎ Π±Π΅Π»ΠΊΠ°, ΠΈΠ½ΠΈΡΠΈΠΈΡΡΡΡΠ΅Π³ΠΎ ΡΠ΅ΠΏΠ»ΠΈΠΊΠ°ΡΠΈΡ Π½ΠΎΠ²ΠΎΠΉ ΠΏΠ»Π°Π·ΠΌΠΈΠ΄Ρ pNB2 ΠΈΠ· Π±Π°ΠΊΡΠ΅ΡΠΈΠΉ Clostridium thermosaccharolyticum.
Π²ΡΠ²ΠΎΠ΄Ρ.
1. ΠΡΠΎΠ²Π΅Π΄Π΅Π½Π½ΡΠΉ ΡΡΠ°Π²Π½ΠΈΡΠ΅Π»ΡΠ½ΡΠΉ Π°Π½Π°Π»ΠΈΠ· Π°ΠΌΠΈΠ½ΠΎΠΊΠΈΡΠ»ΠΎΡΠ½ΠΎΠΉ ΠΏΠΎΡΠ»Π΅Π΄ΠΎΠ²Π°ΡΠ΅Π»ΡΠ½ΠΎΡΡΠΈ RepN Π±Π΅Π»ΠΊΠ° Π²ΡΡΠ²ΠΈΠ» ΠΏΡΡΡ ΠΊΠΎΠ½ΡΠ΅ΡΠ²Π°ΡΠΈΠ²Π½ΡΡ Π΄ΠΎΠΌΠ΅Π½ΠΎΠ², Ρ Π°ΡΠ°ΠΊΡΠ΅ΡΠ½ΡΡ Π΄Π»Ρ Rep Π±Π΅Π»ΠΊΠΎΠ² ΠΏΠ»Π°Π·ΠΌΠΈΠ΄ ΡΠ΅ΠΌΠ΅ΠΉΡΡΠ²Π° pC194/pUBl 10.
2. ΠΠ°ΡΠ΅Π³ΠΈΡΡΡΠΈΡΠΎΠ²Π°Π½Π° ΠΎΠ΄Π½ΠΎΠ½ΠΈΡΠ΅Π²Π°Ρ ΡΠΎΡΠΌΠ° ΠΏΠ»Π°Π·ΠΌΠΈΠ΄Ρ pNB2 Π² ΠΊΠ»Π΅ΡΠΊΠ°Ρ Clostridium thermosaccharolyticum, ΡΡΠΎ ΡΠ²ΠΈΠ΄Π΅ΡΠ΅Π»ΡΡΡΠ²ΡΠ΅Ρ ΠΎ Π΅Π΅ ΡΠ΅ΠΏΠ»ΠΈΠΊΠ°ΡΠΈΠΈ ΠΏΠΎ ΠΌΠ΅Ρ Π°Π½ΠΈΠ·ΠΌΡ ΠΊΠ°ΡΡΡΠ΅ΠΉΡΡ ΠΊΠ°ΡΡΡΠΊΠΈ.
3. ΠΠ΄Π΅Π½ΡΠΈΡΠΈΡΠΈΡΠΎΠ²Π°Π½ ΠΏΡΠΎΠ΄ΡΠΊΡ repN Π³Π΅Π½Π° Π² Π±Π΅ΡΠΊΠ»Π΅ΡΠΎΡΠ½ΠΎΠΉ ΡΠΈΡΡΠ΅ΠΌΠ΅ ΡΡΠ°Π½ΡΠΊΡΠΈΠΏΡΠΈΠΈ-ΡΡΠ°Π½ΡΠ»ΡΡΠΈΠΈ S30 E. coli in vitro.
4. ΠΠ°ΡΠ΅Π³ΠΈΡΡΡΠΈΡΠΎΠ²Π°Π½Π° ΡΠΊΡΠΏΡΠ΅ΡΡΠΈΡ RepN Π±Π΅Π»ΠΊΠ° Π² ΠΊΠ»Π΅ΡΠΊΠ°Ρ E. coli in vivo Ρ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠ΅ΠΌ ΡΡΡΠ΅ΠΊΡΠΈΠ²Π½ΠΎΠΉ ΡΠΈΡΡΠ΅ΠΌΡ Π ΠΠ-ΠΏΠΎΠ»ΠΈΠΌΠ΅ΡΠ°Π·Ρ ΡΠ°Π³Π° Π’7.
5. ΠΡΡΠ°ΡΠΈΠΎΠ½Π½ΡΠΉ Π°Π½Π°Π»ΠΈΠ· ΠΏΠΎΡΠ΅Π½ΡΠΈΠ°Π»ΡΠ½ΡΡ Π°ΠΊΡΠΈΠ²Π½ΡΡ ΡΠ΅Π½ΡΡΠΎΠ² Π±Π΅Π»ΠΊΠ° RepN (Y85 °F ΠΈ Y211F) ΠΏΠΎΠΊΠ°Π·Π°Π», ΡΡΠΎ Π»Π΅ΡΠ°Π»ΡΠ½ΠΎΠ΅ Π΄Π΅ΠΉΡΡΠ²ΠΈΠ΅ Π½Π΅ ΡΠ²ΡΠ·Π°Π½ΠΎ Ρ Π΅Π³ΠΎ Π±Π°Π·ΠΎΠ²ΠΎΠΉ ΡΠΎΠΏΠΎΠΈΠ·ΠΎΠΌΠ΅ΡΠ°Π·Π½ΠΎΠΉ (ΡΠ΅Π»Π°ΠΊΡΠ°Π·Π½ΠΎΠΉ) Π°ΠΊΡΠΈΠ²Π½ΠΎΡΡΡΡ. ΠΠ΅Π»Π΅ΡΠΈΠΎΠ½Π½ΠΎΠ΅ ΠΊΠ°ΡΡΠΈΡΠΎΠ²Π°Π½ΠΈΠ΅ ΠΎΠ±Π»Π°ΡΡΠΈ RepN Π±Π΅Π»ΠΊΠ°, ΠΎΡΠ²Π΅ΡΡΡΠ²Π΅Π½Π½ΠΎΠΉ Π·Π° Π΅Π³ΠΎ Π»Π΅ΡΠ°Π»ΡΠ½ΠΎΠ΅ Π΄Π΅ΠΉΡΡΠ²ΠΈΠ΅, ΠΏΠΎΠΊΠ°Π·Π°Π»ΠΎ, ΡΡΠΎ ΡΡΡ ΡΡΠ½ΠΊΡΠΈΡ ΠΌΠΎΠ³ΡΡ Π½Π΅Π·Π°Π²ΠΈΡΠΈΠΌΠΎ Π΄ΡΡΠ³ ΠΎΡ Π΄ΡΡΠ³Π° ΠΎΡΡΡΠ΅ΡΡΠ²Π»ΡΡΡ Π΅Π³ΠΎ NΠΈ Π‘ — ΠΊΠΎΠ½ΡΠ΅Π²ΡΠ΅ ΡΠ°ΡΡΠΈ, ΠΊΠ°ΠΆΠ΄Π°Ρ ΠΈΠ· ΠΊΠΎΡΠΎΡΡΡ ΡΠΎΠ΄Π΅ΡΠΆΠΈΡ ΠΏΡΠΈΠΌΠ΅ΡΠ½ΠΎ 30% ΠΎΡ ΠΎΠ±ΡΠ΅ΠΉ Π΄Π»ΠΈΠ½Ρ ΠΌΠΎΠ»Π΅ΠΊΡΠ»Ρ.
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- Π ΠΈΡ. 24. Π Π΅ΡΡΡΠΈΠΊΡΠΈΠΎΠ½Π½ΡΠ΅ ΠΊΠ°ΡΡΡ ΠΏΠ»Π°Π·ΠΌΠΈΠ΄ pBluescript II (ΠΏΡΠΎΠ΄ΠΎΠ»ΠΆΠ΅Π½ΠΈΠ΅).1. Esp3 I 4027
- Dra I 3988 Msc I 3980 Sty I Nco I
- Ssp I 3935. Sea I 3830 BstB I 3716 Tthlll X 3698 Drd 136 951. EcoRI 1
- BspM II 5 Dra I 83 I PuuII 103
- Bsv36 13 591 Bel I 3541 BstB I 3716 BsfY I 3440 BspM II 343 Bsg I 34 231. Beg I 32 961. BsaA I 310 Esp3 I 324
- Pvu II 515 Nhel 583 Ace I 594 Xcal 594 Xmn 1635 Bsg I 665
- HincII 321K Drd Π 3175 JSco57 I 3150″ BsaA 13 141
- PpuM I 2974 Msc I 2938, PpuM I 2932, Ava I 2919 Sty I 28 631. Bgl I 26 571. BspM I 255
- Nru I 246″ Eag I 2433 Bgl I 24 231. Beg I 2202 Hinc II 2145
- BspH I 1983 Sph I Nsp I 20 561. Sac Π 8321. Π’ΠΠΠ II 8721. V 1679
- Nhel 1723 BamH I BsfY I 1869 Ban II 1979,1965
- BstB I 1318 Β¦Ase I 1405 Ssp I 1418 Xbal 1424 'Ear I 1446 ' Eco57 I 1463 Tthlll II 1501 Clal 1517 β’ Hind Π¨ 1523
- Ace I 2145 IECONI2U6 Sail 2145
- Π ΠΈΡ. 24. Π Π΅ΡΡΡΠΈΠΊΡΠΈΠΎΠ½Π½Π°Ρ ΠΊΠ°ΡΡΠ° pACYC184.1. See 142 991. Aatlt 47 411. Siyl 1824
- Π ΠΈΡ. 25. Π‘ΡΡΡΠΊΡΡΡΠ° Π²Π΅ΠΊΡΠΎΡΠ° pALTER-1.
- EBSSmHX? P S. P S.m Π XHSmSBE1. KIXX1. Kan'1. EBSaP1. Sm H1. PSaBE1. Π Β¦= EcoR 11. Π = BamHI1. S = Sacl1. Sm β’=Sm"l1. H «= Hind III1. X β’=Xhol1. P -=Pstt1. Sp =Spft 11. Sa «Sa/11. Π «Kpnl1. A = ΠΡΠ°1
- Π ΠΈΡ. 27. Π Π΅ΡΡΡΠΈΠΊΠ΄ΠΈΠΎΠ½Π½ΡΠ΅ ΠΊΠ°ΡΡΡ ΠΏΠ»Π°Π·ΠΌΠΈΠ΄ pUC4K ΠΈ pUC4KIXX.
- Π’7{ TncTag WicI Hti’Tij thnxrtxnsita S"Tag ΠΠ« Π Kpnlenterofcinase site1. Wool1. EccRVmti1. EecRI1. Sac I1. Sal I1. HnJII1. Wofl1. Eifl-Ixnol1. Aval1. Htj-Tag0pot1O2l1. T7lemiinalor
- Π ΠΈΡ. 28. Π Π΅ΡΡΡΠΈΠΊΡΠΈΠΎΠ½Π½Π°Ρ ΠΊΠ°ΡΡΠ° ΠΏΠ»Π°Π·ΠΌΠΈΠ΄Ρ ΡΠΠ’32.