Cloning of C-P Compound Biodegrading Genes in Pseudomonas sp. strain #A1

Pseudomonas sp. strain #A1에서 C-P 화합물 분해 유전자의 Cloning

C-P compounds(Pn; phosphonate) such as glyphosate(GPS), aminoethylphosphonate(AEPn) and methyl-phosphonate(MPn) biodegrading genes were cloned from Pseudomonas sp. strain #A1 Which assimilated GPS as sole phosphorous source. Carrying out the in vivo molecular cloning by means of Mini-Mu plasmid, the size of clones($AEPn^+$, $MPn^+$, $GPS^+$) for the gene to degrade C=P compounds are 10-19Kb, 10Kb, and 12-18 Kb, respectively. Moreover, they expressed the phenotype for each Pn when they were transformed into $\Delta phn$ mutants. Hence, it is postulated that Pseudomonas sp.#A1 has three kind of Pn degradative pathway, separately. The phn clones($AEPn^+$, $MPn^+$, $GPS^+$) are verified as the members of PHO regulon because of their phoBR-dependent characteristics.

C-P 화합물(Pn; phosphonate)의 일종인 glyphosate(GPS)를 인산원으로 이용하는 Pseudomonas sp. strain #A1으로부터 GPS 분해 유전자 및 2-aminoethylphosphonate(AEPn), methyl-phosphonate(MPn)와 같은 Pn의 분해 유전자를 클로닝하였다. Mini-Mu plasmid를 이용한 in vivo molecular cloning 결과 약 10-19 Kb의 $AEPn^+$ clones, 10 Kb의 $MPn^+$ clones, 12-18 Kb의 $GPS^+$ clone들을 얻었으며 E. coli의 $\Delta phn$ mutants에 transformation 하였을 때 각각의 Pn에 대한 다양한 phenotype을 나타냈다. 따라서 Pseudomonas sp. strain #A1에서는 적어도 3종류의 Pn 분해대사 경로를 갖고 있는 것으로 예측된다. 뿐만 아니라, 각각의 phn clone($AEPn^+$, $MPn^+$, $GPS^+$)들은 PHO regulon의 조절유전자 phoBR에 의존하여 발현하였다.