Restriction digests of the clone give the following sizes (kb): BamHI--6.0; BglII--6.0; HindIII--6.0. The glucocerebrosidase insert contains the following restriction sites (approximate kb from the 5' end): HindIII--0.83; PstI--0.27, 0.56; SalI--1.25; BamHI--1.48. Contains the complete coding sequence for glucocerebrosidase (nt 6-1556), plus 290 nt at the 3' end. Corresponds approximately to nt 490-2336 of the sequence record, with a modification at the 5' end from CCCACATGG to CCACCATGG. The vector was derived from pSV2-dhfr by adding the adenovirus promoter and removing the poison sequence from the pBR322 portion. Permits bicistronic expression, with glucocerebrosidase being translated before dhfr, resulting in greater expression of the insert sequence relative to dhfr. The order of the major features in the plasmid is: pMB1 ori - SV40 enhancer - Adenovirus major late promoter - XhoI - BglII - glucocerebrosidase insert - dihydrofolate reductase - SV40 polyadenylation - ampR. |
