
¡Ø º» Á¦Ç°Àº CloningÀ» À§ÇØ In-Fusion
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Á¦Ç° »ç¿ë Àü In-Fusion¢ç ¿ø¸®(µ¿¿µ»ó¸µÅ©)¸¦ ¹Ýµå½Ã Âü°íÇØÁֽñ⠹ٶø´Ï´Ù.
[ÁÖÀÇ] µ¿°áµÈ ¼¼Æ÷ÁÖÀÇ Çص¿(Thawing) ½Ã, ¼¼Æ÷ »ýÁ¸À² Çâ»óÀ» À§ÇÏ¿© collagen-coated plate ¶Ç´Â flaks¸¦ »ç¿ëÇϽʽÿÀ.
ÃßÈÄ ¼¼Æ÷ ¹è¾ç ½Ã¿¡µµ ¼¼Æ÷ÁÖÀÇ ºÎÂøÀ²(adherence)ÀÌ ÁÁÁö ¾Ê´Ù¸é collagen-coated plate
¶Ç´Â flask¸¦ »ç¿ëÇÏ´Â °ÍÀ» ±ÇÀåÇÕ´Ï´Ù.
NOTE: We recommend using collagen-coated plates or flasks for efficient culturing of frozen stocks. Vessels coated
with compounds other than collagen may also provide suitable growth substrates (e.g. poly-L-lysine), but only collagen-coated plates
(e.g. BD BioCoat Cellware, Collagen Type I) have been tested at Clontech. Once recovered, the cells may be cultured directly on tissue
culture plastic. However, if adherence is poor, we recommend using only collagen-coated vessels.
- °í¿ª°¡ÀÇ ÀçÁ¶ÇÕ retroivirus¸¦ È¿°úÀûÀÌ°í ½Å¼ÓÇÏ°Ô »ý»ê
- ´Ù¾çÇÑ ¼¼Æ÷¿¡ °¨¿°
Retro-XTM Universal Packaging System
Retro-X Universal Packaging SystemÀº GP2-293 packaging cell lines°ú ¸ñÀûÀ¸·Î ÇÏ´Â cell line¿¡ µû¶ó packaged virus°¡ ģȼºÀ» º¸À̵µ·Ï ÇÏ´Â 4 °³ÀÇ envelope vector (pVSV-G, pEco, pAmpho, p101A1)·Î ±¸¼ºµÇ¾î ÀÖÀ¸¸ç, °í¿ª°¡ÀÇ amphotropic, ecotropic, dualtropic ¹× pantropic virus¸¦ 48 ½Ã°£ À̳»¿¡ ¾òÀ» ¼ö ÀÖ´Ù. ÀÌ packaging systemÀº Pantropic Expression System°ú °°Àº cell lines¿¡ »ç¿ëÇÑ´Ù.
Pantropic Retroviral Expression System
Pantropic Retroviral Expression SystemÀº VSV-G virus envelope glyco proteinÀ» ¹ßÇöÇÏ´Â ¹ÙÀÌ·¯½º¸¦ »ý»êÇϱâ À§ÇÏ¿© GP2-293 cell lineÀ» »ç¿ëÇÑ´Ù. ÀÌ glycoproteinÀº ¼¼Æ÷ Ç¥¸éÀÇ receptor¿¡ ÀÇÁ¸ÀûÀÌÁö´Â ¾ÊÁö¸¸ lipid °áÇÕ ÇÏ¿© plasma membrane°úÀÇ À¶ÇÕÇÏ¿© ¹ÙÀÌ·¯½º°¡ ħÅõÇÏ´Â °ÍÀ» ¸Å°³ÇÑ´Ù (2).
RetroPackTM PT67 cell lines
RetroPack PT67Àº 10A1 viral envelop¸¦ ¹ßÇö½ÃÅ°´Â NIH3T3 ±â¹ÝÀÇ packaging cell lineÀÌ´Ù. PT67 ¼¼Æ÷¿¡¼ packagingµÈ ¹ÙÀÌ·¯½º´Â ´Ù¾çÇÑ Æ÷À¯·ù ¼¼Æ÷¸¦ °¨¿°½ÃÅ°´Âµ¥ »ç¿ëµÈ´Ù. ¿Ö³ÄÇÏ¸é ¹ÙÀÌ·¯½º°¡ µÎ °³ÀÇ ´Ù¸¥ Ç¥¸é ºÐÀÚÀÎ amphotropic retroivirus receptor (Ram-1)¿Í GALVr receptor¸¦ ÅëÇØ ¼¼Æ÷³»·Î µé¾î°¥ ¼ö Àֱ⠶§¹®ÀÌ´Ù. RetroPack PT67 Cell LineÀº Retro-X system¿¡ Æ÷ÇԵǾî ÀÖ´Ù.
Retroivirus packaging cell line°ú systemÀº ClontechÀÇ ¸ðµç retroivirus vector¿Í ÇÔ²² »ç¿ëÇÒ ¼ö ÀÖ´Ù.
Table I: Retroviral Packaging Cell Lines
Integrated Markers |
Product |
Cell Type |
Tropism |
Envelope |
Receptors |
gag-pol |
env |
Host Cell |
RetroPack PT67 Cell Line |
NIH 3T3 |
Dualtropic |
10A1 |
GALVr, Ram-1 |
TK |
DHFR |
Many mammalian cell types |
GP2-293 Cell Line* |
HEK 293 |
Pantropic |
** |
** |
** |
n/a |
All cell types |
* Retro-X Universal Packaging System and the Pantropic Retroviral Expression System.
** The GP2-293 Cell Line requires cotransfection with one of several types of envelope protein vectors and can produce virus with various tropisms.
Components & Storage Conditions
°¢ Á¦Ç° ±¸¼º¹°°ú º¸°üÁ¶°ÇÀº Certificate of Analysis ¸¦ ÂüÁ¶ÇϽʽÿÀ.
References
1. Burns, J. C. et al. (1993) Proc. Natl. Acad. Sci. USA 90:8033-8037.
2. Emi, N. et al. (1991) J. Virol. 65:1202-1207.