The insert contains 2 copies of a 2.2 kb insert containing the following restriction sites, separated by approximate kb: EcoRI- 0.4- HindIII- 0.1- BglII- 0.6- PvuII- 0.2- BglII- 0.9- EcoRI. Restriction digests of the clone give the following sizes (kb): EcoRI--2.7, 2.4, 2.4; HindIII--4.55, 2.4, 0.55; PvuII--2.4, 2.4, 1.4, 1.3; BamHI--7.5; HindIII--7.5. The EcoRI RFLP shows significant linkage to Wiskott-Aldrich Syndrome (theta = 0.00, Z = 10.19). The 0.9 BglII/EcoRI fragment contains a 334 bp HaeIII/Sau3A fragment within which a deletion of about 3 kb occurred during cloning. The 334 bp HaeIII/Sau3A fragment contains 3 complete copies and a partial copy of a 26 bp repeat which includes a 10 bp perfect inverted repeat separated by 3 nt. EcoRI gives the best separation of alleles on 0.8% agarose gels. A stringency of 0.5 X SSC, 64 C was used routinely. |
