เอกสารการประชุมวิชาการและเสนอผลงานวิจัย มหาวิทยาลัยทักษิณ ครั้งที่ 19 2552 - page 926

3
„µ¦ž¦³»¤ª·µ„µ¦Â¨³Áœ°Ÿ¨Šµœª·‹´¥¤®µª·š¥µ¨´¥š´„¬·– ‡¦´Ê Šš¸É 19 ž¦³‹Îµže 2552
°»ž„¦–r¨³ª·›¸„µ¦
1. ®¤´„Áž¨º°„´ž³¦——o ª¥°´˜¦µn ªœ °ŠÁž¨º°„´ž³¦— (g) ˜n°œÊ嘵¨ (g) ‡º° 3:1 Áž} œÁª¨µ 12 ª´œ
×¥«¹„¬µž¦·¤µ–Þ¦˜¸œ ¨³ž¦·¤µ– °ŠÁ°œÅŽ¤rÁŽ¨¨¼Á¨Â¨³Ãž¦˜· Á°
2. œÎµœÊ室´„Áž¨º°„´ž³¦— ¤µšÎµ„µ¦Â¥„Á°œÅŽ¤rÁŽ¨¨¼Á¨Â¨³Ãž¦˜·Á°—oª¥Áš‡œ·‡ Aqueous two-
phase system ץčo Polyethylene glycol 1500 (PEG 1500) š¸É‡ªµ¤Á o¤ oœ 5%, 10%, 15% ¨³ 20% w/v ¨³Äo
Á„¨º° 3 œ·— ŗo„n Tripotassium citrate, Ammonium sulphate ¨³ Dipotassium hydrogen phosphate ¨oªÁ ¸¥œ
„¦µ¢ Binomial curve Á¡ºÉ°®µ´ —nªœ °Š¡°¨·Á¤°¦r¨³Á„¨º°š¸ÉÁ®¤µ³¤ ¨oªÁ¨º°„‡ªµ¤Á o¤ oœ °Š¡°¨·Á¤°¦r¨³
œ·— °ŠÁ„¨º °š¸É µ¤µ¦™Â¥„Á°œÅŽ¤rÁŽ¨¨¼Á¨Â¨³Á°œÅŽ¤rÞ¦˜·Á°Å—o —¸š¸É »—
3. Â¥„Á°œÅŽ¤rÁŽ¨¨¼Á¨Â¨³Ãž¦˜·Á°ÄœœÊ室´„Áž¨º°„´ž³¦—ץčo°´˜¦µnªœ °Š¡°¨·Á¤°¦r¨³
Á„¨º°š¸ÉÁ®¤µ³¤ ¨oªœÎµ´Êœ¡°¨· Á¤°¦r¨³´ÊœÁ„¨º°¤µšÎµ„µ¦„ε‹´—¡°¨· Á¤°¦r¨³Á„¨º°°°„—oª¥Áš‡œ·‡ Gel filtration
ץčo Sepharose 6B œµ—‡°¨´¤œr 30 cm × 2 cm ´¢Á¢°¦rš¸É čo‡º° 50 mM Tris-HCl pH 7.0 ¨³Äo °´˜¦µ„µ¦Å®¨
Ášnµ„´ 0.22 ml/min Á„ȝ Fraction ¨³ 2 ml ¡¦o°¤š´ÊŠ®µž¦· ¤µ–Þ¦˜¸œ—oª¥ª·›¸ Lowry method (1951) ž¦·¤µ–
Á°œÅŽ¤rÁŽ¨¨¼Á¨Â¨³ž¦·¤µ–Á°œÅŽ¤rÞ¦˜·Á°
4. œÎµ Fraction š¸É˜¦ª‹¡Á°œÅŽ¤r˜n¨³œ·—¤µ®µœÊ宜´„äÁ¨„»¨—oª¥Áš‡œ·‡ Sodium dodecyl sulphate
polyacrylamide gel electrophoresis (SDS-PAGE) ץčo 12% Separating gel ¨³ 4% Stacking gel
Ÿ¨„µ¦ª·‹´¥Â¨³°£· ž¦µ¥
‹µ„„µ¦«¹„¬µœÊ室´„®¤´„Áž¨º°„´ž³¦—š¸É®¤´ „Áž}œ¦³¥³Áª¨µ 12 ª´ œ Ĝ£µª³Å¦o°°„Ž·Á‹œ ¡ªnµ ¤¸
ž¦·¤µ–Þ¦˜¸œÁšnµ„´ 13.19 mg/ml ¤¸ž¦·¤µ–Á°œÅŽ¤r ÁŽ¨¨¼Á¨Ášnµ„´ 124.24 Units/ml ¨³¤¸ž¦·¤µ–Á°œÅŽ¤r
Þ¦˜·Á°Ášnµ„´ 11.25 Units/ml ×¥Á°œÅŽ¤rš¸É Á„·— ¹Êœ Á„·—‹µ„„µ¦Ÿ¨·˜ °Š‹»¨·œš¦¸¥rĜ›¦¦¤µ˜·š¸É ¡°¥¼nĜ¦³
„µ¦®¤´ „ ×¥‹»¨·œš¦¸ ¥rÁ®¨nµœ¸Ê‹³Äo Áž¨º°„´ž³¦—Ž¹ÉŠ¤¸š´Ê Š°Š‡rž¦³„°š¸ÉÁž}œÃž¦˜¸ œÂ¨³ÁŽ¨¨¼Ã¨Áž} œÂ®¨nŠ
‡µ¦r °œÁ¡ºÉ°ÄoĜ„µ¦Á‹¦·Á˜·Ã˜
Á¤ºÉ °œÎµœÊ室´„Áž¨º°„´ž³¦—¤µšÎµ„µ¦Â¥„Á°œÅŽ¤r ÁŽ¨¨¼Á¨Â¨³Á°œÅŽ¤r Þ¦˜·Á°—oª¥Áš‡œ· ‡ —oª¥
Áš‡œ·‡ Aqueous two-phase system ץčo Polyethylene glycol 1500 (PEG 1500) š¸É ‡ªµ¤Á o¤ oœ 5%, 10%, 15%
¨³ 20% w/v ¨³ÄoÁ„¨º° 3 œ· — ŗo„n Tripotassium citrate, Ammonium sulphate ¨³ Dipotassium hydrogen
phosphate ‹³Å—o „¦µ¢ Binomial curve —´Š£µ¡š¸É 1 Á¤ºÉ °œÎµž¦·¤µ–Á°œÅŽ¤r ÁŽ¨¨¼Á¨Â¨³Á°œÅŽ¤rÞ¦˜·Á°Äœ´Êœ
µ¦¨³¨µ¥¡°¨·Á¤°¦r (PEG) ¤µ®µ¦—oª¥ž¦·¤µ–Á°œÅŽ¤rÁŽ¨¨¼Á¨Â¨³Á°œÅŽ¤rÞ¦˜·Á°Äœ´Ê œµ¦¨³¨µ¥Á„¨º° ‹³Å—o
‡nµ K Ž¹ÉŠµ¤µ¦™°„Å—oªnµÁ°œÅŽ¤r˜n¨³œ·—Â¥„Åž°¥¼nĜ´Ê œ¡°¨·Á¤°¦r®¦º°Á„¨º°¤µ„„ªnµ„´œ Ÿ¨—´Š„¨nµªÅ—o—Š
Ūo—´Š˜µ¦µŠš¸É 1 ‹µ„Ÿ¨„µ¦«¹„¬µÂ—ŠÄ®oÁ®Èœ „µ¦ÄoÁ„¨º° Tripotassium citrate (K-citrate) ¨³ Dipotassium
hydrogen phosphate (K 2HPO4) Ĝ„µ¦ Patition ‹³Å—o‡nµ %Yield °ŠÁ°œÅŽ¤r ÁŽ¨¨¼Á¨Â¨³Á°œÅŽ¤rÞ¦˜·Á°˜É優nµ
„µ¦ Partition —o ª¥Á„¨º° Ammonium sulphate —´ Šœ´Êœ ‹¹ŠÁ¨º°„čo Á„¨º° Ammonium sulphate ((NH4)2SO4)Ĝ„µ¦
Partition ÁœºÉ°Š‹µ„¤¸Ÿ¨˜n°„µ¦šÎµŠµœ °ŠÁ°œÅŽ¤rš´ÊŠ°Šœ·—œo°¥š¸É »— Á¤ºÉ°Å—o œ·— °ŠÁ„¨º°š¸ÉÁ®¤µ³¤Â¨o ª‹³˜o°Š
¡· ‹µ¦–µ™¹Š´—nªœ °Š¡°¨·Á¤°¦r¨³ž¦· ¤µ–Á„¨º°š¸ÉÁ®¤µ³¤°¸„—oª¥ ×¥‹³¡·‹µ¦–µ‹µ„‡ªµ¤µ¤µ¦™Äœ„µ¦Â¥„
Á°œÅŽ¤r ÁŽ¨¨¼Á¨Â¨³Ãž¦˜·Á°Ž¹ÉŠ‹µ„˜µ¦µŠš¸É 1 ‹³Á®È œÅ—oªnµ „µ¦Äo PEG 15% ¨³Á„¨º° Ammonium sulphate
19.2% ‹³µ¤µ¦™Â¥„Á°œÅŽ¤rÞ¦˜·Á°Â¨³Á°œÅŽ¤rÁŽ¨¨¼Á¨°°„‹µ„„´ œÅ—o—¸š¸É »— ×¥Á°œÅŽ¤rÞ¦˜·Á°‹³Â¥„