The insert contains the following sites separated by (bp)(approx): EcoRI- 340- BglII- 50- BglII- 210- EcoRV- 305- PstI- 920- EcoRI. Restriction digests of the clone give the following sizes (kb): EcoRI--3.0, 2.1; BamHI--5.0; HincII--3.2, 1.4 + smaller; BglII--5.2; XbaI--5.2. The insert contains a 774 bp open reading frame which codes for amino acids 244-501 of human liver GDH, and about 1 kb of untranslated sequence. The cDNA sequence of human brainstem glutamate dehydrogenase is 95% homologous to the human liver sequence and 97% homologous to the bovine liver sequence. This probe detects a single 3.7 kb band in human brain and glioma mRNA. In rat it detects a 3.5 kb band in liver, brain and C6 glioma cells, a 2.8 kb band in kidney, and a reduced 2.8 kb band in brain total mRNA. This clone encodes conserved epitopes because it was isolated from an expression library by anti-bovine glutamate dehydrogenase antibodies. |
