Study On The Interaction Of P6.9Protein In Baculovirus AcMNPV

Concentrating and packing virus DNA into the capsid is essential for baculovirus replication. P6.9protein coded by baculovirus is responsible for enrichment of DNA. Reports show that all the baculoviruses contain P6.9or its homologous proteins, which sizes of those proteins are between49-109amino acids. All P6.9homologous proteins contain function domain. As the Molecular weight of P6.9coded by AcMNPV is6.9kDa, the protein is named as P6.9.This theme chooses AcMNPV P6.9as the research object to explore those proteins which interaction with P6.9protein and further clarify the function of P6.9protein.Chapter1is a general introduction of the background of baculovirus, including baculovirus taxonomy, baculoviral replication and baculovirus proteomics. Additional, two methods for functional analysis of proteins were introduced.Chapter2is to verify the P6.9protein polymerization in vitro. Three fusion proteins labeled different markers were expressed by protokaryon expression system. Those fusion proteins were analyzed in soluble and selected two more soluble to made pulldown experiments. The results revealed the interaction between P6.9on itself, indicating P6.9oligomerization.Chapter3the cDNA library for yeast two-hybrid was constructed. The P6.9was used as a bait to screen the cDNA library. The total RNA was extracted from SF9cells infected by AcMNPV.The RNA was used as template for the construction of yeast-two hybrid cDNA library using the technology of homologous recombination. The AcMNPV SF9cDNA library was detected. The results indicated that the cDNA library was qualified for the yeast two hybrid screening. The bait plasmids pGBK-p6.9was successfully constructed by fusing the encoding fragments of P6.9to the vector pGBKT7. The results of the toxicity and self activating effect showed that suitable to yeast two-hybrid. Then the cDNA library was screened by the P6.9as a bait to identify protein (s) that interacted with P6.9and26candidate positive clones was obtained. BLAST results showed that13cDNAs was P6.9itself, and the others are in sequencing.This chapter also indicated that PK1have no interaction with p6.9by the method of Co-transformation with pGADT7-PK1and pGBKT7-P6.9.Chapter4Summarized and discussed in this article.