full2010.pdf - page 1120

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‡ªµ¤µ¤µ¦™Äœ„µ¦‡Š°¥¼n°Š¡¨µ¤·—¨¼„Ÿ¤ÄœÁºÊ° Escherichia coli 2 µ¥¡´œ›»r
Segregrational Stability of Recombinant Plasmids in Two Different Escherichia coli Strains
‹»”µ¤µ« Á‹¦·» 1 ¨³¤–”¨ Á¨·«‡–µªœ·„»¨1*
Jutamas Jaroensuk 1 and Monthon Lertcanawanichakul1*
š‡´—¥n°
ž{®µÎµ‡´°¥nµŠ®œ¹ÉŠÄœ„µ¦šÎµ¡´œ›»ª·«„¦¦¤‡º°¡¨µ¤·—¨¼„Ÿ¤Å¤n µ¤µ¦™‡Š°¥¼n ŗoĜĜÁŽ¨¨rÁ‹o µoµœ
Ž¹ÉŠÄœ„µ¦š—¨°Šœ¸Êŗo Á¨º°„čo¡¨µ¤·— 4 œ·—nŠÁž}œ 1) ¡¨µ¤·—¡µ®³ 2 œ·—„¨nµª‡º ° pHY300PLK (4.87 „·Ã¨-
Á) ¨³ pBluescript II KS (2.96 „·Ã¨Á) 2) ¡¨µ¤·—¨¼ „Ÿ¤¥¸œÅ‡˜·Áœ 2 œ·—„¨nµª‡º° pCHIB1 (5.54 „· è-
Á) ¨³ pHYB43 (7.45 „·Ã¨Á) Á¡ºÉ°«¹„¬µ‡ªµ¤µ¤µ¦™Äœ„µ¦‡Š°¥¼n°Š¡¨µ¤·——´ Š„¨nµªÄœÁºÊ° Escherichia
coli 2 µ¥¡´œ›»r (DH5v ¨³ INVvF c) Á¤ºÉ°Á¨¸Ê ¥ŠÄœ°µ®µ¦Á¨¸Ê¥ŠÁºÊ ° Luria-Bertani (LB) š¸ÉŤn¤¸¥µž’·¸ªœ³Áž}œ
Áª¨µ 30 ª´œ Ÿ¨ž¦µ„‘ªnµ¡¨µ¤·— œµ—Á¨È„µ¤µ¦™‡Š°¥¼nŗo ĜÁŽ¨¨rÁž}œ¦³¥³¥µªœµœ„ªnµ¡¨µ¤·— œµ—Ä®n¨³
Á¤ºÉ°¤¸ „µ¦œÎµ·Êœ¥¸œÅ‡˜· Áœ°—ÄnÁ oµÅžÄœ¡¨µ¤·—¡µ®³‹³šÎµÄ®o‡ªµ¤µ¤µ¦™Äœ„µ¦‡Š°¥¼n°Š¡¨µ¤·—£µ¥Äœ
ÁŽ¨¨r¨—¨ŠŽ¹ÉŠµ¤µ¦™Á¦¸¥Š¨Îµ—´Å—o—´Šœ¸Ê pHYB43 < pHY300PLK ®¦º° pCHIB1 < pBluescript II KS.
‡ÎµÎµ‡´ : „µ¦‡Š°¥¼n°Š¡¨µ¤·— ¡¨µ¤· —¨¼„Ÿ¤ Ҙ·Áœ
Abstract
One of many problems of genetic engineering technique is the segregrational instability of recombinant
plasmids in host cells. Such, this experiment was set to examine for segregrational stability of recombinant
plasmids in two different Escherichia coli strains (DH5vand INVvFc) . The plasmids were divided into two
groups; one was the plasmid vectors, i.e., pHY300PLK (4.87 kilobase) and pBluescript II KS (2.96 kilobase);
and, the other was recombinant chitinase gene plasmids, i.e., pCHIB1 (5.54 kilobase ) ¨³ pHYB43 (7.45
kilobase). The transformants were separately grown, daily transferred in Luria-Bertani (LB) medium without
antibiotic addition (non selective pressure) for 30 days. The results showed that a small plasmid was more
structurally stable than a large one. Moreover, the inserted-chitinase gene could be effected on the segregrational
stabillty , i.e., the recombinant chitinase gene plasmids were lower segregrationally stable than plasmid vectors as
follow; pHYB43< pHY300PLK or pCHIB1< pBluescript II KS.
Keywords : Segregrational Stability, Recombinant Plasmid, Chitinase
1 µ µÁš‡œ·‡„µ¦Â¡š¥r εœ´„ª·µ®Áª«µ˜¦r¨³µ›µ¦–»«µ˜¦r ¤®µª· š¥µ¨´¥ª¨´ ¥¨´„¬–r œ‡¦«¦¸›¦¦¤¦µ 80160.
1Programme in Medical Tachnology Institute of Allied Health Sciences and Public Health Walailak University Nakhon-Si-
Thammarat 80160.
Corresponding author : Ú¦«´¡šr/Ú¦µ¦ 075672104 / 075672106 E-mail:lmonthon@wu.ac.th