| Comments |
The insert contains the following restriction sites (nt position according to published sequence): PvuII--140, 851, 2506, 2989, 5287; SacI--874, 3548; PstI--1158, 2286; EcoRI--2088, 2377, 2656, 3396, 5018; BamHI--2601, 5975, 6014; BglI--5266. The insert does not contain HindIII or SalI sites. Restriction digests of the clone give the following sizes (kb): EcoRI--5.0, 2.2, 1.9, 1.7, 0.8, 0.3; SacI--4.2, 3.8, 2.8, 0.7, 0.4; PstI--10.7, 1.2; BamHI--3.4 (doublet), 2.9, 1.9, 0.3; BglI--6.6, 4.0, 1.1, 0.2. The coding sequence extends from nt 91 to nt 6765, with the signal peptide encoded from nt 91 to nt 174. May be transformed into bacterial hosts such as E. coli HB101 or DH5, or transfected into CHO or COS cells. The vector contains dihydrofolate reductase coding sequences to permit amplification by methotrexate. Detects a mRNA of about 7.0 kb in liver and HepG2 cells. A SalI-cleavable linker was inserted into the vector EcoRI site and the insert was cloned into that SalI site. The vector contains the SV40 ori, the adenovirus major late promoter and tripartite leader, the SV40 polyadenylation signal, the adenovirus VA I and VA II genes, and pBR322 sequences. |
