Abstract

The conjugally transferred TOL plasmid or NAH plasmid was stably maintained and expressed in n-alkane assimilating Pseudomonas putida KCTC 2405. However, these plasmids were not able to coexist in this strain because of incompatibility. The incompatibility of TOL and NAH plasmid was bypassed using CAM::TOL* plasmid, which was constructed by the transposition of only tol gene without incompatibility system in TOL plasmid into CAM plasmid. p. putida 3SK capable of growing on m-toluate, naphthalene, camphor, and n-alkane(C8-C24) was constructed by the conjugal transfer of NAH plasmid into n-alkane assimilating p. putida SK carrying CAM:: TOL* plasmid. CAM::TOL＊ plasmid in p. putida 3SK was stable on the selective media but unstable on the nonselective media.

TOL 플라스미드와 NAH 플라스미드는 n-알칸을 자화하는 P. putida KCTC 2405에 접합에 의해 각각의 이동은 가능하나 두 플라스미드는 불화합성에 기인하여 본 균주내에 공존할 수 없었다. TOL plasmid에서 불화합성 체계는 남겨두고 tol 유전자만 이 CAM plasmid내로 transposition 되어 형성된 CAM::TOL* 플라스미드는 NAH 플라스미드와 P. putida KCTC 2405에서 공존할 수 있어 m-toluate, naphthalene, camphor 및 n-alkane(C8-C24)를 분해할 수 있는 P. putida 3SK 균주를 육종하였다. CAM::TOL* 플라스미드는 선택성 배지에서 안정하였으나 비선택성 배지에서는 불안정하였다.

Keywords

• Conjugal Transfer;
• n-Alkane Assimilation;
• Pseudomonas putida;
• plasmid