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GenBank
|
Entry Name |
Accession No. |
pUC18 |
SYNPUC18CV |
L09136 |
pUC19 |
SYNPUC19CV |
L09137 |
pHSG299 |
SYNHSG299 |
M19415 |
pHSG399 |
SYNHSG399 |
M19087* |
1. pHSG298 DNA (Code 3298)´Â MCS¼¿ ÀÌ¿Ü¿¡´Â pHSG299 DNA¿Í °°´Ù. ´Ù¸¸, 1,446 À§Ä¡ÀÇ ¡®G¡¯°¡ °á¼ÕµÇ¾î ÀÖ´Ù. pHSG299 DNA (Code 3299)´Â GenBank¿¡
µî·ÏµÇ¾î ÀÖ´Â pHSG299 ¼¿°ú ¾Æ·¡ ¹è¿¿¡ Â÷ÀÌ°¡ ÀÖ´Ù.
1633C, 2667-2668 GA¡æ°á¼Õ 1808C¡æT, 2228-2229 AT¡æTA, 2474G¡æA
2. pHSG396 ¹× pHSG398Àº multi-cloning siteÀÇ ¼¿ ÀÌ¿Ü´Â pHSG399¿Í µ¿ÀÏÇÏ´Ù.
ÁÖ) GenBank¿¡ µî·ÏµÇ¾î ÀÖ´Â pUC18¼¿Àº 1308¹øÀÌ GÀÌÁö¸¸, ´ç»ç¿¡¼ ÆǸÅÁßÀÎ pUC18 º¤ÅÍ´Â 1308¹øÀÌ AÀÌ´Ù.
DNA ±æÀÌ
pUC18/pUC19 |
: 2,686 bp |
pHSG298, pHSG299 |
: 2,676 bp |
pHSG396 |
: 2,238 bp |
pHSG398 |
: 2,227 bp |
Á¦Ç°¼³¸í
pUC18/pUC19´Â dideoxy¹ý¿¡ ÀÇÇÑ DNA ¿°±â¼¿ °áÁ¤¿¡ ÀûÇÕÇÑ plasmid vector·Î ampicillin ³»¼ºÀ» °¡Á® M13 phage vector¿¡ ºñ±³ÇÏ¿© Å« DNA ´ÜÆíÀ» cloningÇÒ ¼ö ÀÖ´Ù.
lacZ ¿µ¿ª¿¡ multi-cloning site¸¦ °®°í ÀÖ¾î IPTG¿Í X-galÀ» ÇÔÀ¯ÇÏ´Â plate¿¡¼ ¿Ü·¡ DNAÀÇ µµÀÔ À¯¹«¸¦ °£´ÜÈ÷ ÆǺ°ÇÒ ¼ö ÀÖ´Ù. ¶Ç lac promoter¸¦ ÀÌ¿ëÇÑ ¿Ü·¡ À¯ÀüÀÚÀÇ
¹ßÇöµµ °¡´ÉÇÏ´Ù.
DNA sequencing¿¡´Â M13 primers¸¦ »ç¿ëÇÏ´Â °ÍÀÌ Æí¸®ÇÏ´Ù. ¶Ç Kilo-Sequence¿ë Deletion Kit (Code 6030)¸¦ »ç¿ëÇÏ¿© kilo-sequencingµµ °¡´ÉÇÏ´Ù.
pHSG298°ú pHSG299´Â kanamycin ³»¼ºÀ», pHSG396, pHSG398Àº chloramphenicol ³»¼ºÀ» °¢°¢ÀÇ ¼±Åà ¸¶Ä¿·Î °®´Â pUC type cloning vector plasmidÀÌ´Ù.
pHSG298°ú pHSG398Àº pUC18°ú, pHSG299´Â pUC19¿Í µ¿ÀÏÇÑ multi-cloning site¸¦ °®°í ÀÖ´Ù (´Ü, pHSG298°ú pHSG299´Â Hind III, Sma I ºÎÀ§¸¦ »ç¿ëÇÒ ¼ö ¾ø´Ù).
¶Ç pHSG396Àº pHSG398°ú ´Ù¸¥ multi-cloning site¸¦ °¡Áö°í ÀÖ´Ù.
¿ëµµ
- ¿Ü·¡ À¯ÀüÀÚÀÇ ¥á- »óº¸¼ºÀ» ÀÌ¿ëÇÑ Å¬·Î´×°ú lac promoter¸¦ ÀÌ¿ëÇÑ À¯ÀüÀÚ ¹ßÇö
- M13 primers¸¦ ÀÌ¿ëÇÑ DNA ¿°±â¼¿ °áÁ¤
- Kilo-Sequence¿ë Deletion Kit (TaKaRa Code 6030)¸¦ ÀÌ¿ëÇÑ ±ä ´ÜÆíÀÇ ¿°±â¼¿ °áÁ¤
»ç¿ë»óÀÇ ÁÖÀÇ
pHSG298°ú pHSG299 ÁßÀÇ Stu¥°Àνļ¿Àº Stu¥°À¸·Î Àý´ÜÇÒ ¼ö ¾ø´Ù. ¼¿¿¡´Â ¹®Á¦°¡ ¾ø±â ¶§¹®¿¡ ±¸Á¶»óÀÇ ¹®Á¦·Î »ý°¢µÈ´Ù.
°ü·ÃÁ¦Ç°
M13 primers (Code 3810, 3820, 3830A, 3830B, 3831, 3832A, 3832B)
±×¸². pUC18/ pUC19 DNAÀÇ ±¸Á¶