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ÀûÀýÇÑ foldingÀ» À§ÇÑ chaperone team

Chaperone Plasmid Set

-

Á¦Á¶»ç Á¦Ç°ÄÚµå Á¦Ç°¸í ¿ë·® °¡°Ý
(ºÎ°¡¼¼º°µµ)
ºñ°í »ç¿ëÀڸŴº¾ó
Takara
3340
Chaperone Plasmid Set
°ü·ÃÇмú ±¸¸ÅÇϱâ
1 Kit
312,000¿ø 
390,000¿ø
°¡°ÝÇÒÀÎ
09.02 ~ 10.31
3340_e.v2012Da.pdf

*º» Á¦Ç°±ºÀ» ÀÌ¿ëÇØ »ý»êÇÑ ÀçÁ¶ÇÕ ´Ü¹éÁúÀ» »ó¾÷ÀûÀ¸·Î È°¿ëÇÏ°íÀÚ ÇÏ´Â °æ¿ì, ´ÙÄ«¶óÄÚ¸®¾Æ °í°´Áö¿ø¼¾ÅÍ (02-2081-2510)·Î º°µµ ¹®Àǹٶø´Ï´Ù.

Á¦Ç°¼³¸í
´ëÀå±Õ¿¡¼­ ÀçÁ¶ÇմܹéÁúÀ» ¹ßÇö½ÃÅ°´Â °æ¿ì, ¹ßÇöµÈ ´Ü¹éÁúÀÌ inclusion body¸¦ Çü¼ºÇϰųª protease¿¡ ÀÇÇØ ºÐÇصǴ °æ¿ì°¡ ºó¹øÇÏ°Ô ¹ß»ýÇÑ´Ù. ÀÌ´Â ¹ßÇöµÈ ´Ü¹éÁúÀÌ Á¤È®ÇÏ°Ô folding µÇÁö ¾Ê¾ÒÀ» ¶§ ³ªÅ¸³ª´Â °ÍÀ¸·Î ´Ü¹éÁúÀÇ ±â´É ¿¬±¸¿¡ Å« °É¸²µ¹ÀÌ µÇ°í ÀÖ´Ù. ChaperoneÀ» ÀÌ¿ëÇÏ¸é ´Ü¹éÁúÀÇ Á¤È®ÇÑ folding È¿À²À» ³ôÀÏ ¼ö ÀÖ¾î, in vitro¿¡¼­ ´Ü¹éÁú folding ±âÀÛÀ» ±Ô¸íÇϱâ À§ÇÑ ¸¹Àº ¿¬±¸¿¡ È°¿ëµÇ°í ÀÖ´Ù.
º» Chaperone Plasmid Set´Â ¼­·Î ´Ù¸¥ 5Á¾ÀÇ plasmid·Î ±¸¼ºµÇ¾î ÀÖ°í, °¢ plasmid´Â ÃÖÀûÀÇ ´Ü¹éÁú ¹ßÇö ¹× foldingÀ» °¡´ÉÇÏ°Ô ÇÏ°í ´Ü¹éÁúÀÇ misfoldingÀ» ÁÙÀ̱â À§ÇØ chaperone team À¸·Î ÇÔ²² ±â´ÉÇÏ´Â ¿©·¯ Á¾ÀÇ chaperone ºÐÀÚ°¡ ¹ßÇöÇϵµ·Ï ¼³°èµÇ¾î ÀÖ´Ù.
°¢ plasmid´Â pACYC º¹Á¦ ±âÁ¡ (ori)°ú chloramphenicol ÀúÇ×¼º À¯ÀüÀÚ (Cmr)¸¦ °¡Áö°í ÀÖ¾î ampicillin ÀúÇ×¼ºÀ» ºÎ¿©ÇÏ´Â ColE1 °èÅëÀÇ plasmid¸¦ Æ÷ÇÔÇÏ´Â ´ëÀå±Õ ¹ßÇö½Ã½ºÅÛ°ú ÇÔ²² »ç¿ëÇÒ ¼ö ÀÖ´Ù. ÇÏÁö¸¸, Cmr À¯ÀüÀÚ¸¦ Æ÷ÇÔÇÑ plasmid, chloramphenicol¿¡ ³»¼ºÀ» °¡Áö´Â ´ëÀå±Õ ¼÷ÁÖ¿Í´Â ÀÌ¿ëÇÒ ¼ö ¾ø´Ù. ¿¹¸¦ µé¾î pET½Ã½ºÅÛ µî¿¡¼­ ÀÌ¿ëµÇ´Â E. coli BL21 (DE3)´Â ¼÷ÁÖ·Î »ç¿ëÇÒ ¼ö ÀÖÁö¸¸, Cmr À¯ÀüÀÚ°¡ Æ÷ÇÔµÈ pLysS ¶Ç´Â pLysE plasmid¿Í pACYC º¹Á¦±âÁ¡À» ¸ðµÎ °¡Áö´Â E. coli BL21(DE3) pLysS ¿Í´Â ÇÔ²² »ç¿ëÇÒ ¼ö ¾ø´Ù.
¸ñÀû ´Ü¹éÁú°ú chaperone ºÐÀÚÀÇ co-expressionÀ» À§Çؼ­´Â chaperone plasmid·Î ÇüÁúÀüȯÇÑ ´ëÀå±ÕÀ» Á¦ÀÛÇÏ°í, ±× ÈÄ ¸ñÀû´Ü¹éÁú ¹ßÇöÀ» À§ÇÑ plasmid¸¦ ÇüÁúÀüȯÇÏ´Â 2´Ü°è ¹æ¹ýÀ» ÃßõÇÑ´Ù. µû¶ó¼­ chaperone plasmid·Î ÇüÁúÀüȯÇÑ ´ëÀå±ÕÀ¸·ÎºÎÅÍ competent cellÀ» Á¶Á¦ÇÏ´Â °ÍÀÌ ÇÊ¿äÇϸç, Chaperone Competent Cell BL21 ½Ã¸®Á ÀÌ¿ëÇÏ¿© ÀÌ °úÁ¤À» »ý·«ÇÒ ¼öµµ ÀÖ´Ù.
±¸¼º

1.

Plasmid pG-KJE8 (10 ng/§¡)

100 §¡

2.

Plasmid pGro7 (10 ng/§¡)

100 §¡

3.

Plasmid pKJE7 (10 ng/§¡)

100 §¡

4.

Plasmid pGTf2 (10 ng/§¡)

100 §¡

5.

Plasmid pTf16 (10 ng/§¡)

100 §¡

º¸Á¸ - 20 ¡É
Chaperone plasmids map

No.

Plasmid

Chaperone

Promoter

Inducer

Resistant Marker

Sequence*

1

pG-KJE8

dnaK-dnaJ-grpE
groES-groEL

araB
Pzt-1

L-Arabinose Tetracyclin

Cm

link

2

pGro7

groES-groEL

araB

L-Arabinose

Cm

link

3

pKJE7

dnaK-dnaJ-grpE

araB

L-Arabinose

Cm

link

4

pG-Tf2

groES-groEL-tig

Pzt-1

Tetracyclin

Cm

link

5

pTf16

tig

araB

L-Arabinose

Cm

link

*2020³â 12¿ù sequence Á¤º¸°¡ ¼öÁ¤µÇ¾ú½À´Ï´Ù.

<ÀÌ¿ëÇÒ ¼ö ÀÖ´Â ¹ßÇö¿ë Çö󽺹̵å>
º» Á¦Ç°¿¡ Æ÷ÇÔµÈ Chaperone plasmid´Â pACYCÀÇ º¹Á¦ ±âÁ¡ ¹× chloramphenicol ³»¼º À¯ÀüÀÚ (Cmr À¯ÀüÀÚ)À» Æ÷ÇÔÇÏ°í ÀÖ¾î, °¡Àå ¸¹ÀÌ »ç¿ëµÇ´Â ColE1 ŸÀÔÀÇ ampicillin ³»¼º À¯ÀüÀÚ¸¦ Æ÷ÇÔÇÑ ¹ßÇö¿ë plasmid ½Ã½ºÅÛ¿¡ Àû¿ëÇÒ ¼ö ÀÖ´Ù. °¢ Chaperone À¯ÀüÀڴ araB ¶Ç´Â Pzt-1 (tet) promoter ÇÏ·ù¿¡ ¹èÄ¡µÇ¾î ÀÖ¾î, ¸ñÀû À¯ÀüÀÚ¸¦ lac µî ´Ù¸¥ promoter ÇÏ·ù¿¡ µÎ¸é ¸ñÀû´Ü¹éÁú°ú chaperone ´Ü¹éÁúÀ» µû·Î ¹ßÇö½Ãų ¼ö ÀÖ°í, °¢ promoter¿¡ ÇÊ¿äÇÑ ÄÄÆ÷³ÍÆ® (araC ¶Ç´Â tetR)Àº plasmid »ó¿¡ ¹èÄ¡µÇ¾î ÀÖ´Ù.
´Ü, chloramphenicol ³»¼º À¯ÀüÀÚ¸¦ Æ÷ÇÔÇÑ ¹ßÇö plasmid ¿Í´Â ÇÔ²² »ç¿ëÇÒ ¼ö ¾ø´Ù.
º» Á¦Ç°Àº ƯÈ÷ Cold shock expression vectorÀÎ pCold™ DNA ½Ã¸®Áî (Code 3360-3364), pCold™ TF DNA (Code 3365), pCold™ ProS2 DNA (Code 3371), pCold™ GST DNA (Code 3372)¿Í °°ÀÌ »ç¿ëÇÏ¸é ¸ñÀû ´Ü¹éÁúÀ» ³ôÀº È¿À²·Î ¹ßÇö½Ãų ¼ö ÀÖ´Ù.
°ü·ÃÁ¦Ç°
- °íÈ¿À² E. coli Cold shock ¹ßÇö½Ã½ºÅÛ : pCold™ Vector ½Ã¸®Áî
- Â÷¼¼´ë Membrane ±â¼úÀÇ His-tagged protein Á¤Á¦ column : Capturem™ His-Tagged Purification
- Acetic acid ¾ø´Â °í°¨µµ ´Ü¹éÁú ¿°»ö½Ã¾à : TaKaRa CBB Protein Safe Stain
- ¹ü¿ëÀûÀÎ °ËÃâºÎÅÍ ÃÊ°í°¨µµ °ËÃâ±îÁö ´Ù¾çÇÑ Á¦Ç° ¶óÀξ÷ : HRP Substrate ½Ã¸®Áî

Keyword :

´ëÀå±Õ ´Ü¹éÁú ¹ßÇö
>
ChaperoneÀ» ¹ßÇöÇÏ´Â E. coli BL21 Competent ..
Chaperone Competent Cell BL21 ½Ã¸®Áî
>
ÀûÀýÇÑ foldingÀ» À§ÇÑ chaperone team
Chaperone Plasmid Set
>
GST TagÀÌ Æ÷ÇÔµÈ Cold shock Vector
pCold¢â GST DNA
>
Soluble tagÀÇ °áÁ¤ÆÇ
pCold¢â ProS2 DNA
>
Soluble tag (TF tag) ÀÌ Æ÷ÇÔµÈ Cold shock Ve..
pCold¢â TF DNA
>
°íÈ¿À² E. coli Cold shock ¹ßÇö½Ã½ºÅÛ
pCold¢â Vector ½Ã¸®Áî
>
¸ñÀû ´Ü¹éÁú Àüü¸¦ È¿À²ÀûÀ¸·Î À¯µµ¹ßÇö/µ¿À§..
Single Protein Production System (SPP System¢â)