Cloning and Sequence Analysis of a p40 Structural Protein Gene of Helicoverpa zea Nuclear Polyhedrosis Virus

• Published in: Virology (New York, N.Y.) Vol. 192; no. 1; pp. 224 - 233
• Main Authors: Ma, Shih-Win, Corsaro, Bartholomew G., Klebba, Phillip E., Fraser, M.J.
• Format: Journal Article
• Language: English
• Published: San Diego, CA Elsevier Inc 01.01.1993; Elsevier
• Subjects: Amino Acid Sequence; Antibodies, Monoclonal; Baculoviridae - genetics; Base Sequence; Biological and medical sciences; Cells, Cultured; Cloning, Molecular; DNA, Viral - genetics; Fundamental and applied biological sciences. Psychology; genes; Genes, Viral; Genetics; Helicoverpa zea nuclear polyhedrosis virus; In Vitro Techniques; Microbiology; Molecular Sequence Data; nucleotide sequence; p40 protein; prediction; Protein Structure, Secondary; Restriction Mapping; Sequence Alignment; Sequence Homology, Amino Acid; Solubility; Viral Structural Proteins - genetics; Viral Structural Proteins - immunology; Virology; Entomopathogen; Nucleotide sequence; Insecta; Sequence alignment; Homology; Biosynthesis; Baculovirus; Nuclear polyhedrosis virus; Virus; Proteins; Baculoviridae; Gene; Arthropoda; Lepidoptera; Noctuidae; Kinetics; Invertebrata; Molecular cloning; Aminoacid sequence
• ISSN: 0042-6822; 1096-0341
• DOI: 10.1006/viro.1993.1025

A gene encoding an occluded virion structural protein was isolated from an expression library constructed from the Helicoverpa zea S-type nuclear polyhedrosis virus (NPV) isolate HzS-15 using both polyclonal and monoclonal antibody screening. The gene was located within a PstI-SalI fragment of the HzS-15 genome spanning from 96.5 to 97.3 m.u. Sequencing analyses revealed a long open reading frame of 927 nucleotides that predicted a protein of 37 kDa in size. Immunoblot analyses using the monoclonal antibody ENV409 demonstrated that the gene corresponded to a 40-kDa protein (p40) in SDS-polyacrylamide gels that was present exclusively in enveloped occluded virions but not in extracellular budded virions or envelope-stripped nucleocapsids. A p40 protein-specific transcript was detected at 16 hr postinfection in HzS-15-infected HZ 1075/UND-K cells and remained until 22 hr. Primer extension analyses demonstrated that the p40 protein-specific transcript started at -49 nucleotides from the ATG start codon within an ATAAG consensus pentamer found in late protein genes of Baculoviruses. The deduced amino acid sequence of the HzS-15 p40 protein gene shared 44 and 45% sequence homology with the p40 proteins of Bombyx mori NPV and Autographa californica NPV, respectively.