เอกสารการประชุมวิชาการและเสนอผลงานวิจัย มหาวิทยาลัยทักษิณ ครั้งที่ 19 2552 - page 312

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„µ¦´ŠÁ‡¦µ³®r¦¸‡°¤·ÂœœšrÞ¦˜¸œ Rubber Elongation Factor (HbREF) °Š¥µŠ¡µ¦µ
(Hevea brasiliensis) Ĝ Escherichia coli
Recombinant expression of Hevea Rubber Elongation Factor (HbREF) in Escherichia coli
Áž¨ºÊ°Š » ª¦¦–¤–¸1*, ¤«´„—·Í ͇œ» „¼¨1, œª¨¡¦¦– «·¦·œ» ¡Š«r2, ›œ¡´œ›»r ž{šš¤µœœšr1, ‹µ¦» ª´˜¦ ‹´œš¦rž¦³—·¬“r1,
£´š¦µª»› ‹·ª˜¦³„¼¨3, ¨³ª´ œÁ¡È œ´œšµœ»ª´•œr4
Pluang Suwanmanee1*, Somsak Chocknukul1, Nualpun Sirinupong2, Thanapan Phattamanont1,
Charuvat Chantrapradist1, Patthavut Jewtragoon3, and Wanpen Nuntanuwat 4
š‡´—¥n°
Rubber elongation factor (REF) Áž}œÃž¦˜¸œš¸É¤¸šµšÎµ‡´Äœ„¦³ªœ„µ¦´ŠÁ‡¦µ³®räÁ¨„» ¨¥µŠÄœ
¥µŠ¡µ¦µ ¥¸œ REF (HbREF) ¤¸„µ¦Â—Š°°„Äœ¥µŠ¡µ¦µµ¥¡´œ›»rÄ®oŸ¨Ÿ¨·˜œÊε¥µŠ¼Š¤µ„„ªn µµ¥¡´œ›»rÄ®oŸ¨Ÿ¨· ˜
œÊÎ µ¥µŠ˜Éε ª´˜™» ž¦³Š‡r°ŠŠµœª·‹´ ¥œ¸Ê ˜o°Š„µ¦´ŠÁ‡¦µ³®r¦¸‡°¤·ÂœœšrÞ¦˜¸œ HbREF Ĝ E. coli ץǨœ¥¸œ
HbREF Ĝ pSUMO expression vector ¨³nŠ™nµ¥ (transform) Á oµ¼nÁŽ¨¨r E .coli BL21 plus (DE3) RIL ‡´ —Á¨º°„
Ǩœš¸Éŗo ¦´¦¸‡°¤·Âœœšr¡¨µ¤·— (pSUMO–HbREF ) ˜¦ª‹°˜ÎµÂ®œnŠÂ¨³š·«šµŠ„µ¦ÁºÉ °¤ °Š¥¸œ HbREF
„´ pSUMO vector ×¥ª·›¸ PCR ¨³®µ¨Îµ—´Á ª·Á‡¦µ³®r®µ£µª³š¸É Á®¤µ³¤Äœ„µ¦Á®œ¸É¥ªœÎµÁŽ¨¨rÄ®o
´ ŠÁ‡¦µ³®r¦¸‡°¤·ÂœœšrÞ¦˜¸œ ŗo„n ‡ªµ¤Á o¤ oœ °Š IPTG °» –®£¼¤·Äœ„µ¦n¤ÁŽ¨¨r ¨³¦³¥³Áª¨µÄœ„µ¦
Á®œ¸É¥ªœÎ µÁŽ¨¨r ˜¦ª‹°Ãž¦˜¸œ—oª¥ SDS-PAGE Ÿ¨„µ¦«¹„¬µ¡ªnµµ¤µ¦™‡´ —Á¨º°„Ǩœš¸Éŗo¦´ pSUMO–
HbREF Ž¹ÉŠ¤¸˜ÎµÂ®œnŠÂ¨³š·«šµŠ open reading frame °Š HbREF š¸É™¼„˜o°Šµ¤µ¦™Á®œ¸É¥ªœÎµÁŽ¨¨rÄ®o ´ŠÁ‡¦µ³®r
¦¸‡°¤·ÂœœšrÞ¦˜¸œ °Š HbREF ŗo—¸š¸É¤¸‡ªµ¤Á o¤ oœ IPTG Ášnµ„´ 0.1 mM °» –®£¼¤·n ¤ÁŽ¨¨rÁšnµ„´ 37 qC ¨³
Áª¨µÁ®œ¸É¥ªœÎ µÁŽ¨¨rÁšnµ„´ 6 ´ÉªÃ¤Š ¦¸‡°¤·ÂœœšrÞ¦˜¸œ°¥¼nĜ¦¼ž inclusion body Ĝš»„£µª³š¸ÉÁŽ¨¨r™¼„
Á®œ¸É ¥ªœÎµÂ¨³¤¸„µ¦´ ŠÁ‡¦µ³®rÞ¦˜¸œ
‡ÎµÎµ‡´: rubber elongation factor (REF) ¦¸‡°¤· œœšrÞ¦˜¸œ ¥µŠ¡µ¦µ
1 °µ‹µ¦¥r£µ‡ª·µ¸ªª· š¥µ ‡–³ª·š¥µ«µ˜¦r ¤®µª· š¥µ¨´¥š´„¬·– Š ¨µ 90000
2 Postdoctural Biochemistry and Molecular Biology Wayne State University School of Medicine,
540 E. Canfield Avenue Detroit, MI 48201, United States
3 ™µ´œª·‹´¥¥µŠ „¦¤ª·µ„µ¦Á„¬˜¦ °.®µ—Ä®n ‹.Š ¨µ 90110
4 Ÿ¼onª¥ª·‹´¥ £µ‡ª·µ¸ªª·š¥µ ‡–³ª·š¥µ«µ˜¦r ¤®µª·š¥µ¨´¥š´„¬·– Š ¨µ 90000
*corresponding e-mail:suwanmaneep@yahoo.com