FIG. 4.

In vitro transcription from full-length TGEV constructs. The A-B1, B2-C, and DE-1–F contigs were ligated in vitro as described in the text. (A and B) DNA positions after 8 and 30 h of electrophoresis, respectively. Lane 1, purified A-B1 product; lane 2, purified B2-C product; lane 3, purified DE-1–F product; lane 4, 1-kb ladder; lane 5, in vitro-ligated products; lane 6, high-molecular-weight markers. (C) The in vitro-ligated products were transcribed in vitro, and the products were digested with DNase for 15 min at room temperature and separated in agarose gels. Lane 1, high-molecular-weight DNA markers; lane 2, 1-kb DNA ladder; lane 3, in vitro-ligated TGEV products; lane 4, DNase-treated in vitro transcripts. Arrow indicates high-molecular-weight transcripts.