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Chaperone Æ÷ÇÔ - ´Ü¹éÁú °¡¿ëÈ­ È¿À² Çâ»ó

ProFold Baculovirus Vectors

-

Á¦Á¶»ç Á¦Ç°ÄÚµå Á¦Ç°¸í ¿ë·® °¡°Ý
(ºÎ°¡¼¼º°µµ)
ºñ°í »ç¿ëÀڸŴº¾ó
AB Vector
AV101
ProFold-C1
°ü·ÃÇмú ±¸¸ÅÇϱâ
25 §¡
°¡°Ý¹®ÀÇ  
AB Vector
AV102
ProFold-C2
°ü·ÃÇмú ±¸¸ÅÇϱâ
25 §¡
°¡°Ý¹®ÀÇ  
AB Vector
AV103
ProFold-ER1
°ü·ÃÇмú ±¸¸ÅÇϱâ
25 §¡
°¡°Ý¹®ÀÇ  
AB Vector
AV106
ProFold-PDI
°ü·ÃÇмú ±¸¸ÅÇϱâ
25 §¡
°¡°Ý¹®ÀÇ  
AB Vector
AV100
ProGreen
°ü·ÃÇмú ±¸¸ÅÇϱâ
25 §¡
°¡°Ý¹®ÀÇ  

Á¦Ç° ¼³¸í
ÀçÁ¶ÇÕ baculovirus¸¦ ÀÌ¿ëÇÑ ´Ü¹éÁú ¹ßÇö½Ã½ºÅÛÀº post-translational modification (phosphorylation, glycosylation µî)ÀÌ Æ÷À¯·ù ¼¼Æ÷¿¡ °¡±õ°í, ºÐÀÚ·®ÀÌ Å« ´Ü¹éÁúÀÇ ¹ßÇöÀÌ °¡´ÉÇϸç, °¡¿ë¼º ´Ü¹éÁúÀ» ´ë·® ¹ßÇö½Ãų ¼ö ÀÖ´Â µî E. coli³ª yeast¿Í °°Àº bacteria ½Ã½ºÅÛ¿¡ ºñÇØ ¿©·¯ °¡Áö ÀåÁ¡À» °¡Áö°í ÀÖ¾î Æ÷À¯µ¿¹° À¯·¡ ´Ü¹éÁúÀÇ ´ë·® ¹ßÇö½Ã½ºÅÛÀ¸·Î¼­ ±¸Á¶ Çؼ® ¿¬±¸³ª ±â´É Çؼ® ¿¬±¸¿¡ ÀÀ¿ëµÇ°í ÀÖ´Ù.
±×·¯³ª Á¾Á¾ ¼¼Æ÷ ³»¿¡¼­ °ú¹ßÇö (over-expression)¿¡ ÀÇÇÑ mis-foldingÀ¸·Î ¸ñÀû´Ü¹éÁúÀÌ inclusion body¸¦ Çü¼ºÇϰųª È¿¼Ò¿¡ ÀÇÇØ ºÐÇصǴ °æ¿ì°¡ ¹ß»ýÇÑ´Ù.
ÀÌ·¯ÇÑ Çö»óÀº ´ëºÎºÐÀº ¼¼Æ÷ ³»ÀÇ chaperoneÀÌ °í°¥µÇ¾î ¹ß»ýÇÏ´Â °ÍÀ¸·Î »ý°¢µÇ¾î, chaperone ¹ßÇö vector¸¦ ÇÔ²² µµÀÔÇÏ´Â ¹æ¹ýÀÌ ½ÃµµµÇ°í ÀÖÀ¸³ª, µµÀÔµÈ ¼¼Æ÷ÀÇ chaperone ¹ßÇö·®¿¡ Â÷ÀÌ°¡ ÀÖ¾î ¸ñÀû ´Ü¹éÁúÀÇ °¡¿ë¼º°ú ¹ßÇö ¼öÀ²ÀÇ ¾ÈÁ¤¼º¿¡ ¿µÇâÀ» ¹ÌÄ£´Ù´Â ÇÑ°è°¡ ÀÖ¾ú´Ù (±×¸²1).


±×¸²1. Comparison of multi-vector chaperone systems used in prior studies (A) with ProFold¢â single vector chaperone system (B).

º» ProFold¢â baculovirus DNA´Â ÀÌ·¯ÇÑ ¹®Á¦¸¦ °³¼±Çϱâ À§ÇØ ClontechÀÇ Çٴٰ¢Ã¼º´ ¹ÙÀÌ·¯½º (Autographa californica nuclear polyhedrosis virus; AcNPV) À¯·¡ BacPAK6 Viral DNA (Bsu36¥°digest) (Kitts A.P. and Possee R.D., Biotechniques, 14 :810-817, 1993.)¸¦ °³·®ÇÏ¿© Á¦ÀÛµÈ baculovirus DNAÀÌ´Ù. Çٴٰ¢Ã¼º´ ¹ÙÀÌ·¯½º °¨¿°¸»±â¿¡ ´ë·® ¹ßÇöµÇ´Â polyhedrinÀ» ÄÚµùÇÏ´Â virus genome ºÎÀ§¸¦ Á¦ÇÑÈ¿¼Ò Bsu36 IÀ¸·Î Àý´ÜÇÏ¿© ¼±ÇüÈ­ÇÑ DNA¿¡ Àΰ£ À¯·¡ (ÀϺΠȿ¸ð À¯·¡) chaperoneÀ» ÄÚµùÇÏ´Â À¯ÀüÀÚ¸¦ Æ÷ÇÔÇÏ°í ÀÖ´Ù.
º» Á¦Ç°°ú ¸ñÀûÀ¯ÀüÀÚ¸¦ Æ÷ÇÔÇÏ´Â transfer vector¸¦ °ïÃæ¼¼Æ÷ÁÖ (Sf9, Sf21 µî)¿¡ co-transfectionÇÏ¿© polyhedrinºÎÀ§ÀÇ homologous recombinationÀ» ÅëÇØ ¸ñÀûÀ¯ÀüÀÚ¸¦ ¹ßÇöÇÏ´Â recombinant baculovirus¸¦ È¿À²ÀûÀ¸·Î ¸¸µé ¼ö ÀÖ´Ù. ProFold¢â baculovirus DNA ½Ã¸®Áî´Â ¹ßÇöÇÏ´Â ¸ñÀû ´Ü¹éÁúÀÇ ¼¼Æ÷³» À§Ä¡ (¼¼Æ÷Áú ´Ü¹éÁú, ¸· ´Ü¹éÁú, ºÐºñÇü ´Ü¹éÁú µî)°ú ER (¼ÒÆ÷ü) ·ÎÀÇ À̵¿¼º µîÀ» °í·ÁÇÏ¿© ÃÖÀûÀÇ chaperone Á¶ÇÕÀ» ¼±ÅÃÇÒ ¼ö ÀÖ´Ù (Ç¥1).

Ç¥1. ´Ù¾çÇÑ ProFold¢â baculovirus DNA ½Ã¸®Áî

Marker ¶Ç´Â Chaperone Á¾·ù

GFP

¥â-gal

Hsc70

Hsp40

Calreticulin

PDI

¿ªÇÒ

Áõ½Ä ¸ð´ÏÅ͸µ, ¿ª°¡ÃøÁ¤¿¡ ÀÌ¿ë

Plaque assay¿¡ ÀÌ¿ë

¼¼Æ÷Áú³» Chaperone

¼¼Æ÷Áú³» chaperone

ER ³»°­¿¡¼­ ´ç´Ü¹éÁú folding

ER¿¡¼­ disulfide bond ÀçÁ¶ÇÕ

Á¦Ç°¸í

Code

Protein Expression level

ProGreen¢â

AV100

+++

+++++

 

 

 

 

ProFold-C1¢â

AV101

+++

+++++

+++

++

 

 

ProFold-C2¢â

AV102

+++

++++

+++

+++++

 

 

ProFold-ER1¢â

AV103

+++

+++++

 

 

+++

++++

ProFold-PDI¢â

AV106

 

+++++

 

 

 

++++


ProFold¢â-C1, ProFold¢â-C2´Â ¼¼Æ÷Áú ³»ÀÇ ÁÖ¿äÇÑ chaperoneÀÎ Àΰ£ À¯·¡ Hsc70/Hsp40¸¦ ¹ßÇöÇÏ´Â baculovirus DNA·Î, ¸ñÀû ´Ü¹éÁúÀ» ¼¼Æ÷Áú¿¡¼­ ¹ßÇö½ÃÅ°°íÀÚ ÇÏ´Â °æ¿ì ÃÖÀûÀÌ´Ù.
ProFold¢â-ER1´Â Àΰ£ À¯·¡ calreticulin ¹× PDI (protein disulfide isomerase)°¡ ÄÚµùµÇ¾î ÀÖ°í, ¸·´Ü¹éÁú, ºÐºñ ´Ü¹éÁú, ȤÀº cysteine-rich ´Ü¹éÁú µî ERÀ̵¿¼º ´ç´Ü¹éÁúÀÇ ¹ßÇö¿¡ ÃÖÀûÀÌ´Ù.
¶ÇÇÑ, ProGreen¢â, ProFold¢â-C1, ProFold¢â-C2, ProFold¢â-ER1¿¡´ÂAequorea victoria À¯·¡ÀÇ ³ì»ö Çü±¤ ´Ü¹éÁúÀÎ GFP¸¦ ¸¶Ä¿·Î ¹ßÇöÇϹǷÎ, ¹ÙÀÌ·¯½º Áõ½Ä ¸ð´ÏÅ͸µ, ¹è¾ç Á¶°ÇÀÇ ÃÖÀûÈ­¿¡ À¯¿ëÇÏ°í, À̸¦ control·Î ¿ª°¡È®Àο¡µµ È°¿ëÇÒ ¼ö ÀÖ´Ù.
³»¿ë °¢ baculovirus DNA 25 §¡ (2.5 §¶)
º¸Á¸ 4 ¡É
»ç¿ë»ó ÁÖÀÇ
Á¦Ç° ³»¿¡ »ç¿ë ¸Å´º¾óÀÌ Æ÷ÇԵǾî ÀÖÁö ¾Ê½À´Ï´Ù. Homologous recombination¿¡ µû¸¥ ÀϹÝÀûÀÎ recombinant baculovirus ±¸Ãà ¹æ¹ýÀÌ ÇÊ¿äÇÑ °æ¿ì BacPAK¢â Baculovirus Expression System (Code 631402) Á¦Ç°ÀÇ ¸Å´º¾ó (¸µÅ©)¸¦ Âü°íÇϽñ⠹ٶø´Ï´Ù.
°ü·ÃÁ¦Ç°
- gp64 protein ¿°»öÀ» ÅëÇÑ titration : BacPAK¢â Baculovirus Rapid Titer Kit
- qPCRÀ» ÀÌ¿ëÇÑ ºü¸¥ titration : BacPAK¢â qPCR Titration Kit
- Insect cell line : IPLB-Sf21 Insect Cells
- All-in-One baculovirus system : BacPAK¢â Baculovirus Expression System
- °íÈ¿À² ´Ü¹éÁú ¹ßÇöÀ» À§ÇÑ baculvirus DNA : flashBAC¢â Baculovirus Expression System
- ÃÊ°£´Ü His-Tagged protein Á¤Á¦ Ä÷³ : Capturem¢â His-Tagged Purification ½Ã¸®Áî (mini, maxi, plate µî)

Keyword :

°ïÃæ ¼¼Æ÷ ´Ü¹éÁú ¹ßÇö (Baculovirus)
>
10ºÐ ¿Ï·á - baculovirus ½Å¼Ó¿ª°¡ ÃøÁ¤
BacPAK¢â Baculovirus GoStix¢â Plus
>
°£ÆíÇØÁø baculovirus ¿ª°¡ ÃøÁ¤ Å°Æ®
BacPAK¢â p35 ELISA Kit (Single Wash)
>
Positive / negative control ¿ë baculovirus
Baculovirus control virus / transfer vector
>
Baculovirus DNA & transfer vector set
Baculovirus DNA package kit
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¸é¿ªÇÐÀû ÃøÁ¤¹ýÀ¸·Î 48 ½Ã°£À¸·Î Baculovirus ..
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>
°ïÃæ, Æ÷À¯·ù ¼¼Æ÷¿ë ÀçÁ¶ÇÕ ´Ü¹éÁú °í¹ßÇö ½Ã..
flashBAC¢â Baculovirus Expression System
>
ºÐÀÚ ChaperoneÀ» ¹ßÇöÇÏ´Â Recombinant Baculo..
FoldHelper Helper virus
>
Plaque Assay ¼øÈ­°¡ ºÒÇÊ¿äÇÑ È¿À²ÀûÀÎ Recomb..
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ProFectin
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>
Baculovirus ¿ª°¡£¨Genomic DNA copy ¼ö£©¸¦ ..
qPCR Titer ÃøÁ¤ Kit
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¸ñÀû¿¡ µû¶ó ¼±ÅÃÇÏ¿© Baculovirus Á¦ÀÛ
Transfer Vectors