Rapid sequence determination of late simian virus 40 16S mRNA leader by using inhibitors of reverse transcriptase.

Abstract
A method for the determination of the primary structure of spliced mRNA junction and leader sequences is described. By analogy to the DNA sequencing procedure of Sanger et al., 2'',3''-dideoxynucleoside triphosphates were used as chain-terminating inhibitors of the reverse transcriptase (RNA-dependent DNA polymerase) reaction. By using specific DNA restriction fragments as primers in combination with this technique, the sequence of the spliced junction between the body and the leader sequence of the 16S late mRNA of SV-40 was determined. The method described should be of general utility in mapping spliced mRNA regions for which the corresponding protein sequence (if any) is unknown.