RAPID PCR CLONING AND SEQUENCE DETERMINATION OF THE RAT LITHOSTATHINEGENE

The rat lithostathine gene was isolated from a genomic library using a rapid screening procedure involving PCR amplification. It was charact erized over 2.7 kbp of gene sequence and 2.43 kbp of 5'-flanking seque nce. The 5'-end of the coding sequence was determined by primer extens ion of lithostathine mRNA. The lithostathine sequence spanned over six exons. The promoter region of the gene contained the TATAAA and CCAAT consensus sequences 30 and 107 bp upstream of the cap site, respectiv ely. Furthermore, a tract of (TG)22 repeat, with potential Z-DNA confo rmation, was found at position - 1081.