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. 1999 Jul 6;96(14):7847–7852. doi: 10.1073/pnas.96.14.7847

Figure 2.

Figure 2

Enzymatic activity of the R2 protein mutations. (A) TPRT assay by using a 110-bp 5′ end-labeled target. The two strands of the target DNA are represented by the straight lines, with the labeled 5′ ends noted with an asterisk. The cDNA made in the reaction is indicated by a straight line, and the RNA template is indicated by a dashed line. The 283-nt RNA template contains the sequence of the 3′ untranslated region of the silkmoth R2 element. In the absence of TPRT, 60- and 48-nt labeled DNA fragments are detected on a denaturing gel. If TPRT occurs, a 343-nt fragment also is generated. (B) Autoradiography of the TPRT reaction run on an 8% denaturing polyacrylamide gel. For each reaction, 15 ng (200 fmol) of end-labeled DNA and 4 ng (30 fmol) of protein were incubated for 30 min. Lanes: 1, no protein; 2, wild-type R2 protein; 3, PA..D mutation; 4, PE..D mutation; and 5, YAYD mutation. Numbers to the right indicate the lengths of the observed DNA products.