The insert contains an EcoRI site (1028) and a BglII site (1970). Restriction digests of the clone give the following sizes (kb): HindIII/ClaI--4.3 (doublet), 1.8; EcoRI--4.5, 3.5, 1.1, 0.9; HindIII--6.0, 4.3; BamHI--6.5, 4.0; PstI--3.8, 2.8, 2.0, 1.9. The O.9 kb EcoRI/BglII fragment detects genomic HindIII fragments of the following sizes (kb): 7.0, 1.3, 5.5, 4.5, 7.5, 5.3. The insert consists of the first 3686 nucleotides of the published sequence, including the complete coding sequence (138 - 2922) plus 5' untranslated and partial 3' untranslated regions. The construct expresses a normal-sized protein product in transfected mouse and rat cells. This plasmid was constructed from a 4.3 kb HindIII/ClaI fragment containing 2 MoMLV LTR sequences flanking a beta-lactamase gene, a 2.4 kb SmaI/ClaI fragment containing RSV-promoter-Neo, and the 3.7 kb HindIII/ScaI cDNA fragment. This sequence in the insert was constructed from cDNAs cloned from several sources. |
