Abstract
We sequenced an E. coil 6.6 kb fragment by using in vitro transposition reaction of Tn3. Tn3 inserted randomly into the target plasmid and the ends of the transposon can be used as movable primer sites. Our selection system eliminates cointegrates and leaves only the simple insertions which makes it suitable for sequencing. We applied both the shotgun method and the ordered method in order to compare them. The shotgun method required more clones but produced the total sequence quickly by the use of a suitable software such as DNAMAN. Moreover, the redundancy of sequencing involved in shotgun method has the advantage of making the result more reliable. The ordered method requires information about the order and separation of transposon insertion sites in different clones in order to select the plasmids to be used for sequencing, but resulted in minimum redundancy. However, the amount of preliminary wo rk required in the ordered method before actual sequencing far outweighs the benefit in the reduction in the number of clones necessary to be sequenced. The E. colt: sequence we obtained turned out to be a recombinant between the standard database sequence (AE000441) and another sequence published by Jensen's group (X00781) within the rph gene.
