Restriction digests of the clone give the following sizes (kb): EcoRI/AvaI--2.55, 1.45; PstI--4.0; EcoRI--4.0; AvaI--4.0; HindIII--4.0. The sequence starts at amino acid 363 and continues through a poly(A) tail of 50 bp. The PstI site at the 3' end was not reconstituted. The insert encompasses the last exon including the 3' untranslated region. Codominant segregation for the EcoRI RFLP has been demonstrated in 2 informative families. The gene-related sequence which demonstrates the EcoRI polymorphism maps 15 kb downstream from the alpha-1-antitrypsin gene. Recorded as not polymorphic by Collaborative Research, meaning either not polymorphic in 0.8% agarose gels, or difficult to work with. The clone is for alpha1-antitrypsin but can detect a related sequence. Signals can be distinguished by the sequence and the sizes of fragments generated. At higher stringency this can be made specific for alpha1-antitrypsin. Enzyme(s) not detecting polymorphism (for alpha1-antitrypsin gene-related sequence): BamHI, HindIII, MspI, PstI, XbaI, XhoII (tested against 20 unrelated Caucasians). |
