Restriction digests of the clone give the following sizes (kb): EcoRI--similar to uncut; HindIII--5.8, 0.65; BamHI--6.4; AvaI--4.7, 1.65; SmaI--uncut. The actual cDNA insert is about 600 bp. It cannot be excised as such because of AT tailing, but can be excised as a 1.6 kb MboII fragment containing 845 bp of pMB9. HindIII and MboII patterns of ATCC preparation match that of Forget's isolate. HindIII site 275 bp 3' to AT/exon 1 joint (in exon 2); 0.7 kb HindIII fragment contains 3' end of gene; 5.2 kb fragment contains 5' end; TaqI site 130 bp 5' to HindIII site. The 5' end at the insert coincides exactly with the ATG initiation site, 275 bp from the internal HindIII site. It lacks all the 5' untranslated sequences and the NcoI site bridging the initiation site. |
