Restriction digests of the clone give the following sizes (kb): EcoRI--3.2, 3.0; EcoRI/PvuI--3.2, 1.7, 1.1; PvuII--2.9, 2.5, 0.85; SacI--5.2, 1.3; SacI/EcoRV--3.0, 1.3 (doublet), 0.8; NotI--5.6, 0.8; BamHI--6.6. The insert contains the following restriction sites (bp from the 5' end): EcoRV--2450; NotI--729; PstI--141, 617, 897; PvuII--2661; SacI--1212; SmaI--1340; SphI--499. An RsaI fragment (nt 2014-2224 of the insert, nt 1402-1612 of the published sequence) from intron 4 detects a 15.0 kb HindIII and a 6.4 kb PstI genomic restriction fragment in human DNA, weak HindIII fragments of 4.4 and 3.2 kb in rats, and no hybridizing fragments in Chinese hamsters. Contains sequences beginning 5' to exon to and continuing beyond exon 5. A 1.2 kb SstI/EcoRV fragment (nt 1212-2450 of the insert) detects the following human genomic restriction fragments (kb): COX5BP1-- 3.5 PstI, 15.0 HindIII; COX5BP2--7.9 PstI, 8.4 HindIII; COX5BP3--1.2 PstI, 7.0 HindIII; COX5BP4--3.9 PstI, 4.1 HindIII; COX5BP5--7.6 PstI, 7.0 HindIII; COX5BP6--2.0 PstI, 5.2 HindIII; COX5BP7--6.2 PstI, 16.0 HindIII. Oriented such that the SacI digest gives a restriction fragment of approximately 1.2 kb. Thus the 5' end is adjacent to the T7 promoter. The intact insert can be separated from the vector by a EcoRI/PvuI double digest. |
