Duplication of Tn1000 or gamma-delta begins with the integration of F plasmid via gamma delta sequence into the chromosome of E.coli chromosome forms an unstable Hfr donor(Ra-2). F prime KLF5 (met B+argE+) isolated from such a donor (Ra-2) is equally unstable. When grown in a rich broth KLF5 dissociates into two components, F plasmid and the chromosomal segment. When such an unstable F-prime (KLF5) is introduced into another recA E.coli K-12 recipient already carrying a stable F-prime trp+, the stable recombinant F-prime carrying genetic markers trp +arg+met+ from both F-primes, is formed. Complete physical analysis of such recombinant F-prime plasmid shows the translocation of gamma-delta to a site proximal to phi80att site of F-prime trp plasmid. F component of KLF5 is lost and the lacXYZ operon of F’trp is deleted out! . F plasmid of KLF5 is lost. After these genetic events, the F-prime trp+ plasmid is still temperature sensitive (ts) and has accommodated the chromosomal components of KLF5 stably. In this component the Tn1000 seems to be bonded with metBJF+ argECBH+ operons but their F is lost. Such translocation has occurred in recA strain confirming that Tn1000 together with its chromosomal neighbours (metBJF, argECBH) has become a mobile DNA element.
