Restriction digests of the clone give the following sizes (kb): BamHI/PstI--3.1, 1.0; PstI--3.8; BamHI--3.8. Linearize the plasmid with PstI and use the T7 promoter for sense-strand mRNA synthesis. Linearize with BamHI and use the T3 promoter for antisense-strand synthesis. IL6 has been mapped to the rat chromosome 4. This clone has the same insert as pRIL6C.91 (which was constructed in a modified Okayama and Berg vector). The insert contains nt 13 (XhoI site) through 923 (SspI site) of the full-length rat IL6 cDNA (start ATG is nt 65, stop TAG is nt 698). The mature protein is 187 amino acids, from nt 137 to nt 697. The sequence for this clone is in the GenBank/EMBL data bank with accession numbers M26744 and M26745. The derived rat amino acid sequence is 93% identical to mature murine IL6 and 58% identical to mature human IL6. A 0.9 kb BamHI-PstI fragment of the probe was used to detect a genomic BglI fragment of 13 kb in the rat and genomic BglI fragments of 23 and 6.2 kb in the mouse. |
