Restriction digests of the clone give the following sizes (kb): BamHI--uncut; Sau3AI--1.0, 0.7, 0.6, + smaller; BglII--3.5, 3.2. The sequenced portion of the insert contains the following restriction sites (approximate kb from the 5' end): EcoRV--1.33; HindIII--0.80; NdeI--0.85; PvuII--0.93; EcoRI--1.6. Additional EcoRI sites are known to occur beyond the site indicated. Contains the complete coding sequence (nt 139-1203). A subclone deleting EcoRI fragments downstream of the gene was used for sequencing and expression studies. Proteins of 35, 37 and 38 kDa are expressed. Expression is independent of orientation in the vector, suggesting the presence of an endogenous promoter that functions in E. coli. The vector was constructed from pBluescript SK+ by converting ApaI and SacI sites into BglII sites. The insert in this clone was isolated from ATCC 33958, Elizabethkingia miricola, formerly Flavobacterium meningosepticum. |
