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. 2013 Apr 22;30(7):1653–1664. doi: 10.1093/molbev/mst078

Fig. 1.

Fig. 1.

Assembly, cloning, and expression of polA genes from OCT viral metagenomes. Four reads from the OCT-03 library (panel A, top) were assembled at 75% NAID to form a 7,410-nucleotide consensus contig (panel A, middle) that included three complete and one partial ORF. ORF251 was similar to hel (ACB83959.1), ORF202 to cas4 (ABN70290.1), and ORF ORF1608 had an amino-terminal domain similar to hel (EKQ55894.1) and a carboxy-terminal domain similar to polA (ADC89878.1). Expect values are shown. The sequence immediately adjacent to the 5′-terminus of ORF1608 included consensus transcriptional and translational start elements (panel A, bottom). Primers 2137 and 6768r, shown in panel A, were used to amplify the full-length OCT-1608 ORF from the OCT-07 DNA preparation (panel B). This PCR product (OCT-1608-14) was cloned and expressed to generate a thermostable DNA polymerase and migrates as a 70 kD protein (panel C) seen by SDS-PAGE in the induced (+) but not the uninduced (−) clone. SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis.