항생제에 대하여 다중 저항성을 갖는 Shaphylococcus aµreus D-H-1으로부터 chloram phenicol(Cm) 저항성 유전자를 가진것으로 확인된 R-plasmid(pSBK203, 2.5MdaJ.)를 분리하여 Bacillus subtilis BD 170 내에셔 발현시켰으며 이 Cm 저항성 유전자를 cloning하기 위하여 pSBK203상의 제한효소 인식부위를 결정하였다. pSBK203 상의 TaqI 부분 절편 0.3 kb을 pBD9 CZaI 인식부위내에 삽입하여 얻은 재조합 plasmid(pTQ16)와 TaqI부 분절편과 O.lkb 엇갈럼이 있는 RsaI 단일절떤(1. 3 kb} pBR322의 Seal 인식부위에 삽입하여 얻은 재조합 p plasmid(pHW20)에서 Cm 저항성이 획득되었다. pBD9 및 pBR322 상에 삽입된 두 절펀내에 Hinf, Taq I 빛 BglII의 제한효소 인식부위가 존재하였으며 이들 중 행III 인식부위에 의하여 Cm 저항성이 불활성 되었다.
R-plasmid(pSBK203, 2.5Mdal) conferring chloramphenicol resistance was isolated from mutiple antibiotic resistant Staphylococcus aureus D-H-1. Bacillus subtilis BD170 was transformed by this plasmid and restriction enzyme clevage sites of this plasmid were mapped for the cloning of chloramphenicol resistance gene. Taq I partial digested fragment of pSBK203(1.3kb) inserted into Cla I site of pBD9 appears to have both regulatory region for induction and structural gene for chloramphenicol resistance whereas Rsa I fragment (1.3kb, both ends are staggered away 0.1Kb from those of Taq I fragment) inserted into Sca I site of pBR322 showed constitutive expression in E. coli. Hinf I, Taq I, and Bgl II restriction enzyme recognition sites are found in both Rsa I fragment and Taq I fragment. Among these, Bgl II recognition site was associated with chloramphenicol resistance.
