Fig. 3.

Identification of transcripts and temporal expression analysis of ld109 in the Ld652Y cells infected with LdMNPV. (A) Nucleotide and deduced amino acid sequences of ld109 transcripts and Ld109 protein. Total RNA was extracted from LdMNPV-infected Ld652Y cells at 48 h postinfection by using an RNeasy minikit. RLM-RACE for the ld109 gene was performed by using a FirstChoice RLM-RACE kit. The suggested start (ATG) and stop codons (TAA) are shown by boldface. The transcriptional initiation site (−12) is indicated. The CAGT motifs are boxed by a solid line, and the TATA box-like sequence TAAATAA is underlined. The solid and open arrows indicate the regions used for the preparation of primers for dsRNAs and qRT-PCR analyses, respectively. (B) Temporal analysis of ld109 transcripts. Ld652Y cells were infected with LdMNPV at a multiplicity of infection of 1, and RNA was extracted at the indicated times postinfection by using a Power SYBR green Cells-to-CT kit. These RNAs were treated with DNase I using an RNAqueous-4PCR kit (Ambion) and subjected to reverse transcription. The qRT-PCR analyses of ld109 were performed by using specific primers for ld109 as indicated in panel A. (C) Temporal analysis of Ld109 protein. Polypeptides from LdMNPV-infected Ld652Y cells were analyzed at the indicated times postinfection, and Ld109 protein was probed with the anti-Ld109 antibody. The Ld109 proteins are highlighted by the white dots. For details, see the legend to Fig. 2A.