Development of a <i>Penaeus monodon</i>‐type baculovirus (MBV) DNA probe by polymerase chain reaction and sequence analysis

Abstract

Abstract. Penaeus monodon ‐type baculovirus (MBV) was isolated and purified from the hepatopancreases of MBV‐infected Penaeus monodon Fabricius. MBV DNA was extracted and used as a template in a polymerase chain reaction (PCR). The primers were chosen from conserved regions of the polyhedrin gene of Autographa californica nuclear polyhedrosis virus (AcNPV). One DNA fragment (674 base pairs) was amplified after PCR. There was a 65% homology between the predicted amino acid sequence of this PCR product with that of the polyhedrin polypeptide of AcNPV. Nucleotide sequence analysis indicated that the amplified DNA is the open reading frame of the MBV polyhedrin gene. This 674 bp DNA fragment was subsequently used as a probe in a dot blot analysis. The probe was able to hybridize with the DNA extracted from the purified MBV and from the MBV‐infected P. monodon , but not from the MBV uninfected P. monodon.