Figure 1

Alignments of marker gene sequences derived from Ao38 cells, and those from reference cell lines and insects. A. Alignment of DNA sequences PCR amplified from the mitochondrial cytochrome oxidase c subunit 1 (COI) gene. Primer pairs LCO1490 (5'-GGTCAACAAATCATAAAGATATTGG-3') and HCO2198 (5'-TAAATCTCAGGGTGACCAAAAAATCA-3') [12] were used for PCR amplification of an approximately 658 bp fragment using a previously described method [5] with minor modifications. Reference DNA samples used for comparisons were derived from T. ni larvae (Cornell strain; kindly provided by Dr. Ping Wang), High Five (Tn-5B1-4) cells, Sf9 cells, and an Ascalapha odorata adult. Species identities were also confirmed by analysis using the BOLD database [3, 4, 13]. For all alignments, "." represents nt sequence identity, and differences are indicated by nt abbreviations (A, C, G, or T). B. Alignment of sequences amplified from the internal transcribed spacers (ITS) of the nuclear ribosomal 18S-5.8S-28S cistron. Primer pairs ITS1-1 (5'-CCCCATAAACGAGGAATTCC-3') and ITS4 (5'-TCCTCCGCTTATTGATATGC-3') [8] were used for PCR amplification. An amplicon with a size of approximately 1500 bp was amplified. Reference DNA samples were the same as those used for analysis of COI (above). C. Alignment of cadherin fragment sequences from T. ni larvae and cell lines. The cadherin fragment sequences were PCR amplified from the indicated sources using PCR primers PW-206 (5'-CGCTTTGATGGTCTCGTTC-3') and PW-261 (5'-GCGCTGCTGGGCTTCCTGT-3') as described previously [9]. A 415 nt sequence alignment is shown. Sequences from two T. ni cell lines (High Five and QB-9-4s) and a T. ni larval sample (Cornell strain, provided by Ping Wang) are aligned with the sequence from Ao38 cells.