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ÀüÁ¦Ç°º¸±â
>
Cloning °ü·Ã
>
¼ö½ÄÈ¿¼Ò£¯Nuclease¡¢Topoisomerase
> BAL 31 Nuclease
BAL 31 Nuclease
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Á¦Ç°¸í
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(ºÎ°¡¼¼º°µµ)
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Takara
2510A
BAL 31 Nuclease
50 U
97,000¿ø
Takara
2510B
BAL 31 Nuclease
250 U
436,500¿ø
³óµµ 1~3 U/§¡
Çü»ó
20 mM
Tris-HCl (pH8.0)
100 mM
NaCl
5 mM
CaCl
2
5 mM
MgCl
2
1 mM
EDTA
50%
Glycerol
º¸Á¸ -20¡É
÷ºÎ Buffer Á¶¼º (5¡¿)
40 mM
Tris-HCl (pH8.0)
1,200 mM
NaCl
24 mM
CaCl
2
24 mM
MgCl
2
2 mM
EDTA
±â¿ø
Alteromonas espejiana BAL31
Á¦Ç°¼³¸í
º» È¿¼Ò´Â ÇØ¾ç¼º ¼¼±Õ A. espejiana BAL31°¡ ±Õü¿Ü·Î »ý»êÇÏ´Â ´ÜÀϰ¡´Ú DNA ƯÀÌÀûÀÎ endonucleaseÀÌÁö¸¸, ´ÜÀϰ¡´Ú DNA°¡ ¾øÀ¸¸é ÀÌÁß°¡´Ú DNA¿¡ ÀÛ¿ëÇÏ¿© ¾ç¸»´ÜÀ¸·ÎºÎÅÍ µ¿½Ã¿¡ ºÐÇØÇÏ´Â 5¡¯¡æ 3¡¯, 3¡¯¡æ 5¡¯ exonuclease Ȱ¼º(trimming Ȱ¼º)À» ³ªÅ¸³½´Ù. ÃÖÁ¾ »ê¹°Àº 5¡¯-P mononucleotideÀÌ´Ù. º» È¿¼Ò´Â ºÐÀÚ·® ¾à 83 kDaÀÇ¡²F¡³Çü°ú ºÐÀÚ·® ¾à 73 kDaÀÇ¡² S¡³ÇüÀÌ Á¸ÀçÇϸç,¡² F¡³ÇüÀÌ exo/endoÀÇ È°¼ºÀÇ ºñ°¡ 10~20¹è ³ô´Ù. µÎ Çü¿¡´Â exonulcease Ȱ¼ºÀÌ »ó´ëÀûÀ¸·Î ´Ù¸¥°Í Àܴ̿ ¹ÝÀÀÁ¶°Ç µîÀÇ Â÷ÀÌ´Â È®ÀεÇÁö ¾Ê´Â´Ù. BAL31Àº FÇüÀ» ÁÖ·ÎÇÏ´Â SÇü nuclease¿ÍÀÇ mixtureÀÌ´Ù.
Ȱ¼ºÀÇ Á¤ÀÇ
¿º¯¼º ¼Û¾ÆÁö Èä¼± DNA¸¦ ±âÁú·Î ÇÏ¿© 30¡É, pH8.0ÀÇ Á¶°Ç¿¡¼ 1ºÐ µ¿¾È 1 §¶ÀÇ °¡¿ë¼º ºÐÇØ¹°À» »ý¼ºÇÏ´Â È¿¼ÒȰ¼ºÀ» 1 UÀ¸·Î ÇÑ´Ù.
Ȱ¼º ÃøÁ¤¿ë ¹ÝÀÀ¾× Á¶¼º
20 mM
Tris-HCl (pH8.0)
600 mM
NaC
12 mM
CaCl
2
12 mM
MgCl
2
1 mM
EDTA
650 §¶/ml
±âÁú DNA
»ç¿ë»óÀÇ ÁÖÀÇ
- ´ÜÀϰ¡´Ú DNA endonuclease Ȱ¼ºÀº ¹ÝÀÀ¿Âµµ¸¦ 30 ¡æ 20¡É·Î ³·Ã߸é 1/10 Á¤µµ±îÁö ¶³¾îÁö³ª, dsDNA exonuclease Ȱ¼ºÀÇ ÀÜÁ¸Àº 1/3 Á¤µµÀÌ´Ù.
- Endonuclease Ȱ¼ºÀº ¿°³óµµ¿¡ ¿µÇâ¹ÞÁö ¾ÊÀ¸³ª exonuclease Ȱ¼ºÀº ¿°³óµµ°¡ ³ôÀ»¼ö·Ï ÀúÇÏÇÑ´Ù.
- Trimming Ȱ¼ºÀÇ ÃÖÀû¿°³óµµ´Â 200 mM NaCl ºÎ±ÙÀ¸·Î À̺¸´Ù ³·À¸¸é ¹«ÀÛÀ§ÀûÀ¸·Î ºÐÇØÇÒ ¼ö ÀÖ´Ù.
- ºÐÇØÈ°¼ºÀº ¿°±â¿¡ ´ëÇÑ ÀÇÁ¸¼ºÀÌ ³ô°í GC-richÀÎ ¿µ¿ª(ƯÈ÷ Sma I site)Àº ºÐÇØµÇ±â ¾î·Á¿ì¹Ç·Î ¸»´ÜÀÇ Á¦ÇÑÈ¿¼Ò site¸¦ ¼±ÅÃÇÒ Çʿ䰡 ÀÖ´Ù.
- ¹ÝÀÀÀº distributive È¿¼Ò ³óµµ¿¡ ÀÇÁ¸¼ºÀÌ ³ô°í Èñ¼®¿¡ ´ëÇØ Á÷¼±Àû »ó°üÀº ³ªÅ¸³»Áö ¾ÊÀ¸¸ç, È¿¼Ò¹ÝÀÀÀÌ ³Ê¹« ºü¸¥ °æ¿ì´Â È¿¼Ò·®À» ÁÙÀÌ´Â °Íº¸´Ù´Â ¹ÝÀÀ ¿Âµµ¸¦ ³·Ãß´Â °ÍÀÌ ¹Ù¶÷Á÷ÇÏ´Ù.
- º» È¿¼Ò¿¡ ÀÇÇØ »ý±ä ¸»´ÜÀÇ ÆòȰÈÀ²Àº 10% Á¤µµ·Î ³·°í, ¶óÀ̰ÔÀÌ¼Ç È¿À²À» ³ôÀ̱â À§Çؼ´Â T4 DNA polymerase¿¡ ÀÇÇÑ ¸»´ÜÀÇ ¼öº¹ÀÌ ÇÊ¿äÇÏ´Ù.
¿ëµµ
- DNA ´ÜÆíÀÇ ¾ç¸»´Ü¿¡¼ÀÇ ºÎÅÍÀÇ ÇÑÁ¤ºÐÇØ (°á½ÇÀÇ Á¦ÀÛ)
(°á½ÇÀÇ ±æÀ̰¡1,000¿°±â Á¤µµ°¡ Àû´çÇÏ´Ù. ±× ¿ÜÀÇ °æ¿ì´Â Exonuclease III¿Í S1 nucleaseÀÇ Á¶ÇÕÀ¸·Î ÇÏ´Â °ÍÀÌ ÁÁ´Ù.)
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Keyword :
¼ö½ÄÈ¿¼Ò£¯Nuclease¡¢Topoisomerase
BAL 31 Nuclease
DNA Topoisomerase I
Cloned
Exonuclease I
Cloned
Exonuclease III
Micrococcal Nuclease
Mung Bean Nuclease
RT-PCR Çʼöǰ
Recombinant DNase I (RNase-free)
Cloned
Ribonuclease H (RNase H)
S1 Nuclease