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TaKaRa
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Cloned

SP6 RNA Polymerase

-

Á¦Á¶»ç Á¦Ç°ÄÚµå Á¦Ç°¸í ¿ë·® °¡°Ý
(ºÎ°¡¼¼º°µµ)
ºñ°í »ç¿ëÀڸŴº¾ó
Takara
2520A
SP6 RNA Polymerase
°ü·ÃÇмú ±¸¸ÅÇϱâ
3,000 U
174,400¿ø 
218,000¿ø
°¡°ÝÇÒÀÎ
03.02 ~ 04.30
2520A_DS.v1901Da.pdf
Takara
2520B
SP6 RNA Polymerase
°ü·ÃÇмú ±¸¸ÅÇϱâ
15,000 U
981,000¿ø 

³óµµ 10~50 U/§¡
Çü»ó
10 mM Potassium Phosphate (pH7.9)
1 mM DTT
0.1 mM EDTA
150 mM NaCl
50% Glycerol
º¸Á¸ -20¡É
÷ºÎ Buffer Á¶¼º (10¡¿)
400 mM Tris-HCl (pH7.5)
60 mM MgCl2
20 mM Spermidine
100 mM DTT [º°Ã·]
0.1% BSA [º°Ã·]
±â¿ø
Escherichia coli carrying the plasmid containing phage SP6 RNA polymerase gene
Á¦Ç°¼³¸í
º» È¿¼Ò´Â 5¡¯ ¡æ 3¡¯RNA polymerase Ȱ¼ºÀ» °®À¸¸ç SP6 ÇÁ·Î¸ðÅ並 Æ÷ÇÔÇÏ´Â ÀÌÁß°¡´Ú DNA¸¦ ÁÖÇüÀ¸·Î ÇÏ¿© NTP¸¦ ±âÁú·Î ÇÁ·Î¸ðÅä ÇÏ·ùÀÇ ´ÜÀϰ¡´Ú DNA¿¡ »óº¸ÀûÀÎ RNA¸¦ ÇÕ¼ºÇÑ´Ù. SP6 ÇÁ·Î¸ðÅä ¼­¿­¿¡ ³ôÀº ƯÀ̼ºÀ» ³ªÅ¸³»°í, ´Ù¸¥ »ý¹° À¯·¡ÀÇ promoter´Â ÀνÄÇÏÁö ¾Ê´Â´Ù.
Ȱ¼ºÀÇ Á¤ÀÇ
37¡É, pH7.5ÀÇ Á¶°Ç¿¡¼­ 1½Ã°£ µ¿¾È 1 nmolÀÇ [3H]GMP¸¦ ±âÁú·Î ÇÏ¿© »êºÒ¿ë¼º ħÀü¹°·Î ÇÕ¼ºÇÏ´Â È¿¼ÒȰ¼ºÀ» 1 UÀ¸·Î ÇÑ´Ù.
Ȱ¼ºÃøÁ¤¿ë ¹ÝÀÀ¾× Á¶¼º
40 mM Tris-HCl (pH7.5)
6 mM MgCl2
10 mM DTT
2 mM Spermidine
°¢ 0.5 mM ATP, UTP, CTP
0.5 mM [3H]GTP
0.01% BSA
1 §¶/100 §¡ pSP65 DNA
¼øµµ
- 500 UÀÇ º» È¿¼Ò¿Í 1 §¶ÀÇ ¥ëDNA-Hind III ºÐÇØ¹°À» 37¡É, 16½Ã°£ ¹ÝÀÀÇÏ¿©µµ DNAÀÇ Àü±â¿µµ¿ ÆÐÅÏ¿¡ º¯È­°¡ ¾ø´Ù. - 500 UÀÇ º» È¿¼Ò¿Í 1 §¶ÀÇ 16S¿Í 23S rRNA¸¦ 37¡É, 16½Ã°£ ¹ÝÀÀÇÏ¿©µµ RNAÀÇ Àü±â¿µµ¿ ÆÐÅÏ¿¡ º¯È­°¡ ¾ø´Ù. - 500 UÀÇ º» È¿¼Ò¿Í 1 §¶ÀÇ closed circular (RF I) pBR322 DNA¸¦ 37¡É, 4½Ã°£ ¹ÝÀÀÇÏ¿©µµ nicking Ȱ¼ºÀÌ ¾ø´Ù. - º» È¿¼Ò´Â SDS Àü±â¿µµ¿¿¡ À־, 95% ÀÌ»óÀÇ ¼øµµ¸¦ ³ªÅ¸³½´Ù.
»ç¿ë»óÀÇ ÁÖÀÇ
- ÃÖÀû pH´Â 7.5, Mg2+¿Í ȯ¿øÁ¦¸¦ ¿ä±¸Çϰí, BSA¿Í spermidineÀÇ Ã·°¡¿¡ ÀÇÇØ Ȱ¼ºÈ­µÈ´Ù.
- »ç¿ë½Ã´Â ÷ºÎ ¹öÆÛ¿¡ ¹Ýµå½Ã DTT¿Í BSA¸¦ ÷°¡ÇÒ °Í. À̵éÀ» ÇÔÀ¯ÇÏÁö ¾ÊÀ¸¸é Ȱ¼ºÈ­µÇÁö ¾Ê´Â °æ¿ì°¡ ÀÖ´Ù.
- ÃÖÀû¿Âµµ´Â 40¡É(37¡ÉÀÇ ¾à 130%)·Î, È¿¼Ò·®Àº 300 U/ml±îÁö, ÁÖÇü·®(template DNA)Àº 100 §¶/ml ±îÁö´Â ÇÕ¼º·®¿¡ ´ëÇØ (+)»ó°ü °ü°è¸¦ ³ªÅ¸³½´Ù.
  ¹ÝÀÀ½Ã°£ÀÌ 1½Ã°£À» ÃʰúÇÏ´Â °æ¿ì´Â 37¡É°¡ ¹Ù¶÷Á÷ÇÏ´Ù.
- ƯÁ¤¿µ¿ª¸¸ÀÇ Àü»çÈ¿À²À» ³ôÀ̱â À§Çؼ­´Â ±× ¿µ¿ªÀÇ ÇÏ·ù¿¡ ÁÖÇü DNA¸¦ Æò Ȱ ¶Ç´Â 5¡¯ µ¹Ã⸻´ÜÇüÀ¸·Î Àý´ÜÇÏ´Â °ÍÀÌ ÁÁ´Ù.
¿ëµµ
- Northern hybridization°ú Southern hybridization¿ëÀÇ Ç¥½Ä RNA probeÀÇ Á¦ÀÛ
- RNA splicing ¹ÝÀÀÀÇ Àü±¸Ã¼ÀÇ Á¦ÀÛ
- Cap analog¸¦ ÇÁ¶óÀ̸ӷΠ»ç¿ëÇÑ capped mRNAÀÇ Á¦ÀÛ

Keyword :