中国生物工程杂志

The baculovirus has been widely used as a pesticide in many countries. However, baculovious has some shortcomings such as the speed and spectrum killing insects was slower and narrower than some chemical pesticides. Therefore, the scientists are making efforts to reconstruct baculovirus in order to get new type biological pesticides. Ecdysteroid UDP-glucosyltransferase, egt gene of baculovirus was responsible for the insect exuviate and an important gene. The paper mainly described how to extract SeMNPV genome DNA.The DNA could be successfully used to amplify the purpose gene by polymerase chain reaction(PCR) using SDS-proteinase K-saturated phenol methods，and then the egt gene was cloned from SeMNPV. The result showed that the positive high pure plasmids could be extracted with Alkali method and digested directly with endonuclease. It was known that ORF of SeMNPV egt was 1572bp long and encodes 523 amino acids through DNAstar software analysis. TATA boxes lies at 48~53 and 68~72bp of the 5'-nonencoding region upstream of a translational start site. Comparing with seven kinds of nuclear polyhedrosis virus and one kind of granulosis virus, it indicated that the SeMNPV egt gene is the highest match the Agrotis setegum nuclear polyhedrosis virus(AsNPV).It has 75% nucleotides and 79% amino acid identity. And some of amino acids which closely related to structure and function are very high conservative. Through egt protein phylogenetic tree analysis, it is known that baculovirus would be divided into two classes, one is NPV; and another is granulovirus. The work can lay the foundations for baculovirus genectic engineering in future.