The PstI site at the 3' end of the insert was destroyed during cloning. The retained PstI site is 750 bp from the EcoRI site. The insert contains a HindIII and a XbaI site. Restriction digests of the clone give the following sizes (kb): PstI--5.55; HindIII--4.5, 1.06; RsaI--2.1, 1.4, 0.86, 0.65, 0.56; XbaI/PstI--4.7, 0.8; BglI--2.9, 2.2, 0.4, 0.2; BglI/PstI--2.2, 1.8, 1.0, 0.4, 0.2. Includes 31 bp of 5' and 216 bp of 3' untranslated sequence, and a complete 1008 bp coding sequence. The polyadenylation signal AATAA is 15 bp upstream from poly(A) site. The XbaI site is about 780 bp from the 5' PstI site. A PstI/XbaI double digest should give a probe containing the 5' non-coding sequences and sequences encoding about the first 250 amino acids. Recorded as not polymorphic by Collaborative Research, meaning either not polymorphic in 0.8% agarose gels, or difficult to work with. Thirty hybridizing bands are detected in genomic blots at 37C, 50% formamide, 5X SSC; 100 bands are detected when screening a genomic library under the same conditions. |
