pYUM1118: a new plasmid vector for generating deletions

Abstract

We have constructed a new cloning vector, pYUM1118, for generating deletions. Recently, the kilo-base sequencing method, which uses exonuclease III (ExoIII) and ss DNA specific nuclease, is widely used as a rapid sequencing method of long DNA fragment (>1000 bp) (1). To employ this method, it is necessary to clone the target DNA into the polylinker site of the vectors such as pUC plasmids or M13mp RFs. However,

Sequence and structural analysis of the rfb (O antigen) gene cluster from a group C1 Salmonella enterica strain

“…This clone did agglutinate with 0 7 antibody. The 14 bp EcoRI-Sac1 polylinker region of pPR1250 was then replaced with the 40 bp EcoRI-Sac1 polylinker of pYUM 1 1 18 (Futo et al, 1989), so that orf5.19 was read out of frame. C600 carrying the resultant plasmid pPR1289 did not agglutinate with the antibody, and as ORFs orf6.17 and orf7.17 have very good Shine-Dalgarno sequences and presumably were expressed we conclude both that orf5.19 cannot be complemented by E. coli K12 and that orf5.19 is functional even in the absence of the first 313 bp.…”