Restriction digests of the clone give the following sizes (kb): EcoRI/BamHI--4.0, 3.2, 1.05; ClaI--8.25; HindIII--4.6, 3.7; PstI--4.0, 2.4, 1.0, 0.6, 0.3; BamHI--8.25. The insert contains the following restriction sites (approximate kb from the 5' end): PstI--0.60, 3.0, 3.4; BamHI--0.72; HindIII--4.3; EcoRI--4.7. With prolonged culture on plates, the poly(A) tail becomes shorter. A full-length cDNA clone, with the coding sequence 140 - 4640 bp from the 5' end. A 4.0 kb fragment containing most of the coding sequence can be obtained by a BamHI/EcoRI double digest. The order of the major features in this plasmid is: HindIII - 273 bp SV40 linker - PstI - GC tail (approx. 15 bp) - 5' end - insert - 3' end - AT tail (approx. 110 bp) - 25 bp SV40 - PvuII - pBR322 fragment, including ampR, of 2295 bp. |
