Figure 6.

Schematic map of the recombinant 175-kDa HARE construct. A 5′-end fragment (1.1 kb) of the 175-kDa HARE open reading frame was amplified by RT-PCR with primers containing EcoRI sites, and cloned in-frame with the Ig κ-chain leader sequence of the pSecTag2 vector. The DNA insert was cut with NheI and AsnI and then cloned into pcDNA3.1 together with two other 175-kDa HARE cDNA fragments of 2.2 and 1.4 kb, which were derived from the RT-PCR and cDNA library screenings, respectively, and which encode the remainder of the 175-kDa HARE protein. The fragments were digested with different restriction enzymes as indicated and assembled as described in MATERIALS AND METHODS.