Schumann W, Bade E G, Delius H, Hübert P
Mol Gen Genet. 1978 Mar 20;160(1):115-8. doi: 10.1007/BF00275127.
The DNA of an E. coli K12 strain harboring ten wildtype Mu prophages was restricted with endonuclease EcoRI, and the fragments ligated into the plasmid vector pMB9. Upon transformation of a strain carrying a heat inducible (Mu cts62) prophage, one temperature-resistant transformant was isolated. This transformant strain harbors the hybrid plasmid pKN001, containing the EcoRI.C fragment of Mu DNA as shown by restriction and heteroduplex analysis. Stable transformants of pKN001 are immune to superinfection with phage Mu. Transformation of Mu sensitive bacteria with pKN001 results in killing of the recipients (10(-4) surviving bacteria). The killing function is not expressed upon transformation of Mu-immune (lysogenic) bacteria.
用核酸内切酶EcoRI切割携带10个野生型Mu原噬菌体的大肠杆菌K12菌株的DNA,并将片段连接到质粒载体pMB9中。将携带热诱导型(Mu cts62)原噬菌体的菌株进行转化后,分离出一个抗温度的转化体。通过限制性内切酶和异源双链分析表明,该转化体菌株含有杂交质粒pKN001,其中包含Mu DNA的EcoRI.C片段。pKN001的稳定转化体对噬菌体Mu的超感染具有免疫性。用pKN001转化对Mu敏感的细菌会导致受体菌死亡(存活细菌为10^(-4))。在对Mu免疫(溶原性)细菌进行转化时,杀伤功能不表达。