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ChaperoneÀ» ¹ßÇöÇÏ´Â E. coli BL21 Competent Cell

Chaperone Competent Cell BL21 ½Ã¸®Áî

-

Á¦Á¶»ç Á¦Ç°ÄÚµå Á¦Ç°¸í ¿ë·® °¡°Ý
(ºÎ°¡¼¼º°µµ)
ºñ°í »ç¿ëÀڸŴº¾ó
Takara
9120
Chaperone Competent Cells BL21 Set
°ü·ÃÇмú ±¸¸ÅÇϱâ LMOÁ¦Ç° 
100 ¥ìl¡¿3¡¿6 Á¾·ù
588,000¿ø 
735,000¿ø
°¡°ÝÇÒÀÎ
09.02 ~ 10.31
9120-9125_e.v2102Da.pdf
Takara
9121
Chaperone Competent Cell pG-KJE8/BL21
°ü·ÃÇмú ±¸¸ÅÇϱâ LMOÁ¦Ç° 
100 ¥ìl¡¿10
292,800¿ø 
366,000¿ø
°¡°ÝÇÒÀÎ
09.02 ~ 10.31
9120-9125_e.v2102Da.pdf
Takara
9122
Chaperone Competent Cell pGro7/BL21
°ü·ÃÇмú ±¸¸ÅÇϱâ LMOÁ¦Ç° 
100 ¥ìl¡¿10
292,800¿ø 
366,000¿ø
°¡°ÝÇÒÀÎ
09.02 ~ 10.31
9120-9125_e.v2102Da.pdf
Takara
9124
Chaperone Competent Cell pG-Tf2/BL21
°ü·ÃÇмú ±¸¸ÅÇϱâ LMOÁ¦Ç° 
100 ¥ìl¡¿10
292,800¿ø 
366,000¿ø
°¡°ÝÇÒÀÎ
09.02 ~ 10.31
9120-9125_e.v2102Da.pdf
Takara
9126
TaKaRa Competent Cell BL21
°ü·ÃÇмú ±¸¸ÅÇϱâ LMOÁ¦Ç° 
100 ¥ìl¡¿10
292,800¿ø 
366,000¿ø
°¡°ÝÇÒÀÎ
09.02 ~ 10.31
9126_e.v2007Da.pdf

Á¦Ç°¼³¸í
º» Á¦Ç°Àº Chaperone Plasmid Set (Code 3340)¿¡ Æ÷ÇԵǾî ÀÖ´Â 5Á¾·ùÀÇ chaperone plasmid¸¦ °¢°¢ ÇüÁú ÀüȯÇÑ E. coli BL21ÁÖÀÇ competent cellÀÌ´Ù.
º» Á¦Ç°¿¡ ÀÖ´Â 5 Á¾·ùÀÇ plasmid (pG-KJE8, pGro7, pKJE7, pG-Tf2, pTf16)´Â ´Ü¹éÁúÀÇ folding¿¡ ÇÔ²² °ü¿©ÇÑ´Ù°í ¾Ë·ÁÁ® ÀÖ´Â ¿©·¯ °¡Áö ºÐÀÚ chaperoneÀÌ °¢°¢ ÇϳªÀÇ plasmid»ó¿¡ ÄÚµå µÇ¾î chaperone ÆÀÀ¸·Î È¿À²ÀûÀ¸·Î ¹ßÇöÇϵµ·Ï ¼³°èµÇ¾î ÀÖ´Ù. ÀÌ·¯ÇÑ Chaperone ÆÀ°ú ¸ñÀû ´Ü¹éÁúÀ» co-expression½ÃÅ°´Â °ÍÀ¸·Î ¹ßÇö ´Ü¹éÁúÀÇ °¡¿ëÈ­¸¦ ÃËÁø½Ãų ¼ö ÀÖ´Ù.
E. coli BL21ÁÖ´Â lon protease, ompT ¿Ü¸· protease°¡ °á¼ÕµÈ BÁÖ À¯·¡ÀÇ ±ÕÁַνá, ¹ßÇö ´Ü¹éÁúÀÇ ¾ÈÁ¤¼ºÀ» Çâ»ó½Ãų ¼ö ÀÖ¾î ÀçÁ¶ÇÕ ´Ü¹éÁú ¹ßÇö¿¡ ³Î¸® ÀÌ¿ëµÇ°í ÀÖ´Ù.
ÀϹÝÀûÀ¸·Î Chaperone plasmid¸¦ ÀÌ¿ëÇÏ¿© ¸ñÀû ´Ü¹éÁú°ú chaperone ÆÀÀÇ co-expressionÀ» ½Ç½ÃÇÏ·Á¸é, ¸ÕÀú chaperone plasmid·Î ¼÷ÁÖ E. coli¸¦ ÇüÁú ÀüȯÇÏ¿© ÇüÁú Àüȯü·Î competent cellÀ» Á¦ÀÛÇÑ ÈÄ, ¸ñÀû ´Ü¹éÁúÀ» ¹ßÇöÇÏ´Â plasmid¸¦ ÇüÁú Àüȯ½ÃÄÑ¾ß ÇÑ´Ù.
º» Á¦Ç°Àº Chaperone plasmid ·Î ÇüÁú ÀüȯÇÑ BL21ÁַκÎÅÍ Á¶Á¦ÇÑ competent cellÀ̹ǷΠÇÑ ¹øÀÇ ÇüÁú ÀüȯÀ¸·Î °£´ÜÇÏ°Ô ¸ñÀû ´Ü¹éÁú°ú chaperone ÆÀÀÌ co-expressionµÈ E. coli¸¦ ¾òÀ» ¼ö ÀÖ´Ù.
Chaperone plasmid 5 Á¾·ù ¿Ü¿¡ chaperone plasmid¸¦ Æ÷ÇÔÇÏÁö ¾Ê¾Æ control¿ëÀ¸·Î »ç¿ëÇÒ ¼ö ÀÖ´Â BL21 competent cellµµ ÆǸÅÇÏ°í ÀÖ´Ù.
º» Á¦Ç°Àº ƯÈ÷ Cold shock expression vector pCold DNA ½Ã¸®Áî (Code 3360-3364)¿Í ÇÔ²² »ç¿ëÇÏ¸é ³ôÀº È¿°ú¸¦ ¾òÀ» ¼ö ÀÖ´Ù.

ÁÖ) º» Á¦Ç°ÀÇ ¼÷ÁÖÀÎ BL21ÁÖ´Â T7 RNA Polymerase¸¦ ¹ßÇöÇÏÁö ¾Ê¾Æ, pET ½Ã½ºÅÛ µî T7 promoter¸¦ ÀÌ¿ëÇÑ ¹ßÇö ½Ã½ºÅÛ¿¡´Â »ç¿ëÇÒ ¼ö ¾ø´Ù.
ÁÖ) º» Á¦Ç°Àº Electroporation¿¡´Â »ç¿ëÇÒ ¼ö ¾ø½À´Ï´Ù.
±¸¼º
Chaperone Competent Cells BL21 Set (Code 9120)

1.

Chaperone Competent Cells pG-KJE8/BL21

100 §¡¡¿ 3°³

2.

Chaperone Competent Cells pGro7/BL21

100 §¡¡¿ 3°³

3.

Chaperone Competent Cells pKJE7/BL21

100 §¡¡¿ 3°³

4.

Chaperone Competent Cells pG-Tf2/BL21

100 §¡¡¿ 3°³

5.

Chaperone Competent Cells pTf16/BL21

100 §¡¡¿ 3°³

6.

TaKaRa Competent Cells BL21

100 §¡¡¿ 3°³

7.

pUC19 DNA (0.1 ng/§¡)

10 §¡¡¿ 1°³

8.

3. SOC Medium*

1 §¢¡¿ 18°³


¤ý Chaperone Competent Cells pG-KJE8/BL21 (Code 9121)
¤ý Chaperone Competent Cells pGro7/BL21 (Code 9122)
¤ý Chaperone Competent Cells pG-Tf2/BL21 (Code 9124)
¤ý Chaperone Competent Cells pTf16/BL21 (Code 9125)
¤ý TaKaRa Competent Cells BL21 (Code 9126)

Chaperone Competent Cell (Code 9121~9126)

1.

TaKaRa Competent Cells BL21

100 §¡¡¿ 10°³

2.

pUC19 DNA (0.1 ng/§¡)

10 §¡¡¿ 1°³

3.

SOC Medium*

1 §¢¡¿ 10°³


* SOC Medium Á¶¼º
2 %

Tryptone

0.5 %

Yeast extract

10 mM

NaCl

2.5 mM

KCl

10 mM

MgSO4

10 mM

MgCl2

20 mM

Glucose


[ÁÖÀÇ]
SOC MediumÀ» ³ìÀÏ ¶§ °£È¤ ħÀüÀÌ »ý±â´Â °æ¿ì°¡ ÀÖ½À´Ï´Ù.
ÀÌ·¯ÇÑ °æ¿ì, ½Ç¿Â¿¡¼­ Àß ¼¯°Å³ª 37¡É¿¡¼­ ¸î ºÐ°£ µ¥¿ö¼­ ħÀü¹°À» ³ìÀÎ ÈÄ »ç¿ëÇϼ¼¿ä.
º¸Á¸
-80 ¡É
Chaperone plasmid 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


<ÀÌ¿ëÇÒ ¼ö ÀÖ´Â ¹ßÇö¿ë Çö󽺹̵å>
º» Á¦Ç°¿¡ Æ÷ÇÔµÈ Chaperone plasmid´Â pACYCÀÇ º¹Á¦ ±âÁ¡ ¹× chrolamphenicol ³»¼º À¯ÀüÀÚ (Cm r À¯ÀüÀÚ)À» Æ÷ÇÔÇÏ°í ÀÖ¾î °¡Àå ³Î¸® »ç¿ëµÇ´Â ColE1 ŸÀÔÀÇ ampicillin ³»¼º À¯ÀüÀÚ¸¦ Æ÷ÇÔÇÑ ¹ßÇö¿ë plasmid¸¦ »ç¿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, Chrolamphenicol 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¢â DNA ½Ã¸®Áî (Code 3360-3364), pCold¢â TF DNA (Code 3365), pCold¢â ProS2 DNA (Code 3371), pCold¢â GST DNA (Code 3372)¿Í °°ÀÌ »ç¿ëÇÏ¸é ¸ñÀû ´Ü¹éÁúÀ» ³ôÀº È¿À²·Î ¹ßÇö½Ãų ¼ö ÀÖ´Ù.
Genotype
E.coli BL21£ºF-ompThsdS B (rB- mB-), galdcm
°ü·ÃÁ¦Ç°
- °íÈ¿À² E. coli Cold shock ¹ßÇö½Ã½ºÅÛ : pCold¢â Vector ½Ã¸®Áî
- Â÷¼¼´ë Membrane ±â¼úÀÇ His-tagged protein Á¤Á¦ column : Capturem¢â 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)