Baculovirus genome sequence download

Oreilly1 department of biology, imperial college, london sw7 2az,1 ecology and biocontrol group, centre for ecology and. A phylogeny based on concatenated baculovirus core gene amino acid sequence alignments placed. The genome was 98,392 bp long, which makes it the smallest lepidopteraninfecting. The genometools genome analysis system is a free collection of bioinformatics tools in the realm of genome informatics combined into a single binary named gt. Pseudoplusia includens single nucleopolyhedrovirus psinsnpvie is a baculovirus recently identified in our laboratory, with high pathogenicity to the soybean looper, chrysodeixis includens lepidoptera. The utility of three independent character sets was assessed. In this report we describe the complete genome sequence and organization of the granulovirus isolate hycugvhc1 that infects the larval stages. Genome sequence of an enhancin generich nucleopolyhedrovirus. In this report we describe the complete genome sequence and organization of the granulovirus. Very recently, the in vitro recombinant hbv baculovirus system has been used to study antiviral drug sensitivity of wildtype and mutant hbv. Of the remaining 11 predicted genes, one is related to a lepidopteran serpin gene.

Baculoviridae baculoviridae dsdna viruses international. A major step forward was the development of bacmid technology the construction of bacterial artificial chromosomes containing the genome of the baculovirus which allows the manipulation of the baculovirus genome in bacteria. Interspersed throughout the genome are sections of repetitive sequences of dna known as homologous regions, or hrs. Marek m, van oers mm, devaraj ff, vlak jm, merten ow. Individual repeats are separated by intervening sequences 01 nucleotides in length fig. Initial experiments injecting viruses containing the escherichia coli lacz gene attached to the hsp promoter into embryos of drosophila, the red flour beetle tribolium, and xenopus laevis. Although acmnpv fails to replicate in vertebrate cells, it does express alien genes with levels of expression that are dependent on the strength. The genome of a baculovirus isolated from hemileuca sp. The genome sequence and evolution of baculoviruses. Gene classes early, late and very late are not clustered on the baculovirus genome, and both strands of the genome are involved in coding functions. To facilitate work with these vectors, we sequenced the e2 genome, compared it to that of the acmnpv c6 strain, and found that they are very similar overall.

A phylogenetic tree of all available baculovirus genomes. The viral genomes resource is a collection of viral genomic sequences that is a part of the entrez genomes, which provides curated sequence data and. Crisprcas9 vectors for genome editing and host engineering. Comparative analysis of the complete genome sequences of baculoviruses revealed a core set of 30 genes, 20 of which have known functions. However, this tool has not yet been adopted for use in the baculovirusinsect cell system, which is an important recombinant protein production platform. The complete genome of a baculovirus isolated from an insect of. Genome sequence of chrysodeixis chalcites nucleopolyhedrovirus, a baculovirus with two dna photolyase genes monique m. Engineering of baculovirus vectors for the manufacture of virionfree biopharmaceuticals. Complete genome sequence of the autographa californica. Later, the utility of baculovirus as gene expression vectors was evidenced and a new research area emerged. Recent evidence from genome sequence analyses demands a substantial revision of the taxonomy and classification of the family baculoviridae. The genome sequence of pseudoplusia includens single. P143 is a helicase that requires atp to unwind dna.

Us20180305717a1 baculovirus system for the expression of a. The genome sequence of a baculovirus from hemileuca sp. Crisprcas9 is a powerful sitespecific genome editing tool that has been used to genetically engineer many different systems. Analysis of the genome of the lymantria dispar mnpv ldmnpv, revealed a single orf ld with predicted signal and transmembrane domains. There are ten unique orfs and they account for 3 % of the genome in total. The genome was 98,392 bp long, which makes it the smallest lepidopteraninfecting baculovirus sequenced to date. Origins of truncated supplementary capsid proteins in raav8. Crambidae, one of the most important insect pests of the sugarcane and other monocot cultures in brazil. Comparison of the baculovirus genomes included in this study revealed that 63 genes are common to these nine genomes table 2. Baculovirus was chosen as a vector because it is relatively safe and easy to use, has a broad host range and has a large capacity for dna inserts. One hundred and fortythree agsenpva orfs are homologues of previously reported baculovirus gene sequences.

Expression vector the knowledge that expression of protein coding gene can be induced by placing this gene downstream of a promoter has led to the development of a number of expression vectors, both prokaryotic as well as eukaryotic. Pyralidae, is a common pest of stored goods with a worldwide distribution. The sequence of dnapol is the most conserved among baculovirus replication lefs. Many vectors that are commonly used in the baculovirus insect cell system bics are derived from the autographa californica multiple nucleopolyhedrovirus acmnpv strain e2. Crisprcas9 is a powerful sitespecific genomeediting tool that has been used to genetically engineer many different systems. The baculoviridae is a large group of insect viruses containing circular doublestranded dna genomes of 80 to 180 kbp. Reconstructing gene orders in the ancestral species. Genomewide survey for genes horizontally transferred from. During the last decade, 29 baculovirus genomes have been sequenced offering a wealth of information on the genetic diversity, gene sequences, gene content. Baculovirus as mammalian cell expression vector for gene. Genometools the versatile open source genome analysis software. The complete genome sequence for a larval pathogen of this moth, the baculovirus plodia interpunctella granulovirus pigv, was determined by nextgeneration sequencing.

Sequence and analysis of the genome of a baculovirus. Sep 01, 2003 generation of recombinant baculovirus by direct cloning into a baculovirus genome is also possible 11,12 as well as the use of the cre. Topics including taxonomy, genome replication, early and. Baculoviruses are known to infect invertebrates, with over 600 host species having been described. All 62 lepidopteran baculovirus genes, including the 29 core baculovirus genes, were found in the agsenpva genome. On the other hand, the sequence in semnpv appears to have originated from a separate transfer of a eukaryotic gene. Use of whole genome sequence data to infer baculovirus phylogeny. The complete genome of a baculovirus isolated from an insect. Relatedness of baculovirus and gypsy retrotransposon envelope. In addition, no homologues of polyhedrin or granulin genes were identified in the genome sequence. The pigv genome was found to be 112, 536 bp in length with a 44. Us20180305717a1 us16029,870 us201816029870a us2018305717a1 us 20180305717 a1 us20180305717 a1 us 20180305717a1 us 201816029870 a us201816029870 a us 201816029870a us 2018305717 a1 us2018305717 a1 us 2018305717a1 authority us united states prior art keywords genome recombinant cell locus baculovirus prior art date 20110727 legal status the legal status is. New multipledeletion method for the corynebacterium glutamicum genome, using a mutant lox sequence. Jun 10, 2016 the complete genome of a baculovirus isolated from an insect of medical interest.

Thirty one core genes with orthologs in all genomes were identified along with other. Pdf the genome sequence and evolution of baculoviruses. Analysis of the genome sequence of an alpaca coronavirus. The phylogenetic tree was based on analysis of the combined sequences of 29 conserved genes found in all sequenced baculovirus genomes. The complete genome sequence is 80000180000 nucleotides long. Baculovirus phylogeny based on genome rearrangements. These major divisions within the family baculoviridae were also. Several baculovirus genome sequences have now been pub lished 1, 2, 8, 20, 23, 25, 30, 35, permitting the use of whole genome approaches to infer their phylogeny. It contains 153 putative, nonoverlapping open reading frames orfs encoding predicted proteins of more than 50 aa, together making up 898 % of the genome. As with the previous editions, this information was updated and then integrated into chapters covering the major processes central to the replication and pathology of baculoviruses. Bestbac dna inside the insect cell, the gene of interest is introduced into the baculovirus genome and orf1629 is restored allowing for replication of only recombinant baculovirus. Aug 22, 2017 crisprcas9 is a powerful sitespecific genomeediting tool that has been used to genetically engineer many different systems. There are currently 76 species in this family, divided among four genera.

The cdna sequencing of the static mixed sample yielded altogether 95,953 sequencing reads mapped to the acmnpv genome strain e2 genebank accession. The sequence contains 151 predicted open reading frames orfs with a minimal size of 50 codons. The genome sequence and evolution of baculoviruses annual. Bevs are viable and can infect susceptible hosts, usually cultured lepidopteran insect cells or larvae, in a. Us10017783b2 us14234,988 us201214234988a us10017783b2 us 10017783 b2 us10017783 b2 us 10017783b2 us 201214234988 a us201214234988 a us 201214234988a us 10017783 b2 us10017783 b2 us 10017783b2 authority us united states prior art keywords genome recombinant cell baculovirus genes prior art date 20110727 legal status the legal status is an assumption and is not a legal conclusion. Characterization of ld, indicated that it localizes to. Bestbac linearized baculovirus dna instructions for use. There are several advantages of using baculovirus expression system over e. Jan 05, 1999 sequence and analysis of the genome of a baculovirus pathogenic for lymantria dispar. Us20180305717a1 baculovirus system for the expression of. One report shows that genetic modification of human mesenchymal stem cells hmscs and tissue engineering are also done by the baculoviral vectors 216. The baculovirus genome is nonsegmented and contains a molecule of circular, doublestranded dna. Sequence and analysis of the genome of a baculovirus pathogenic for lymantria dispar.

The complete genome of a baculovirus isolated from an. Late genes may be further subdivided into late and very late genes. Noveltyofacmnpv basedvector the ability to efficiently transfer eukaryotic genes for reli. The vcath and chia dna region to be eliminated from the acmnpv genome was chosen according to kaba et al. Positions of the restriction sites in relation to the baculovirus genome are marked by short vertical lines, with intervening distances given in kilobases. Because the baculovirus genome is normally considered too large for direct insertion of a foreign gene, the gene of interest is first cloned into a transfer vector containing sequences that flank the polh gene in the viral genome. Additionally, other endogenous viral genes related to a rhabdovirus are also present in the sf9. The complete genome sequence of plodia interpunctella. The complete genome sequence of disagv was determined using the 454pyrosequencing method.

Genome sequence analysis and organization of the hyphantria. A total of 12 unique orfs were identified with no homologs in other sequenced baculovirus genomes. The monopoly of insect cells to host baculovirus autographa californica multiple nuclear polyhedrosis virus acmnpv as a eukaryotic gene expression system has been shattered with the growing evidence that it also infects mammalian cells in culture. The complete genome of a baculovirus isolated from an insect of medical interest.

In particular, autographa californica multicapsid nucleopolyhedovirus acmnpv dis are enriched in an origin of viral dna replication ori not associated with the. Perturbing gene expression with baculovirus genome biology. Us10017783b2 baculovirus system for the expression of a. There are currently 84 species in this family, divided among four genera baculoviruses are known to infect invertebrates, with over 600 host species having been described. Bevs are viable and can infect susceptible hosts, usually cultured lepidopteran insect cells or larvae, in a helperindependent fashion. Thus, we created new crisprcas9 vectors that can be used for genome editing in two relevant insect cell lines. Spontaneous excision of bac vector sequences from bacmid. To construct a consolidated recombinant baculovirus expressing the aav2 rep and aav1 cap proteins from the same baculovirus genome, the 2. The new genome sequence was obtained by first mapping reads to a reference sarscov2 genome using bwamem 0. Baculovirus phylogeny based on complete genome sequence data. Recently a number of complete baculovirus genome sequences have been described and were found to lack an open reading frame with homology to gp64. Perturbing gene expression with baculovirus genome. Repeated baculovirus infections in cultured insect cells lead to the generation of defective interfering viruses dis, which accumulate at the expense of the intact helper virus and compromise heterologous protein expression. Use of whole genome sequence data to infer baculovirus phylogeny elisabeth a.

A naturally occurring baculovirus, hyphantria cunea granulovirus hycugvhc1, isolated from the larvae of h. Transcription of baculovirus genes is temporally regulated, and two main classes of genes are recognized. Nterminal tag fusion sequence that may be utilized if the. However, the sequence did match positions 693 to 709 of cun085, an orf with no homology to any other known baculovirus gene. The similarity of these orfs with those of other completely. For customers who prefer a eukaryotic expression system, are exploring lowercost alternatives to mammalian expression system but. Novel baculovirus dna elements strongly stimulate activities. Generation of recombinant baculovirus by direct cloning into a baculovirus genome is also possible 11,12 as well as the use of the cre. Wholegenome sequence of the spodoptera frugiperda sf9. The sf9 cell line is known to produce lowlevel reverse transcriptase rt activity 1, 2, which might be associated with particles. Complete genome sequence of a 2019 novel coronavirus sars.

The large size of the cun085 product 882 amino acids suggests that it is cleaved to produce components of cuninpv obs. Several phylogenetic methods based on whole genome sequence data were evaluated using data from nine complete baculovirus genomes. This is the 4th edition of a book that was initiated with the annotation of the function of all the genes in the most commonly studied baculovirus, acmnpv. The sequence from slmnpv clustered with animal molecules and in particular those of insects. This number is lower than that previously reported by chen et al.

Spodoptera frugiperda cell lines and their derivatives are used to produce a variety of baculovirusexpressed biological products. A betabaculovirus disagv was isolated from diatraea saccharalis lepidoptera. Baculovirus molecular biology oregon state university. The invention relates to a recombinant baculoviral genome useful for the production of viral vectors for gene therapy, allowing said production from a single infection baculovirus system for the expression of a gene therapy vector. Deletion of vcath and chia from the acmnpv bacmid was performed in the li dh10bac strain containing the acmnpv bacmid that was transformed with the pkd46 plasmid to allow lambda red recombination. The baculovirus polh gene is nonessential for virus replication in insect cell cultures and therefore can be removed from the virus genome with no detrimental effect to bv production 214,215. Mar 17, 2000 baculovirus was chosen as a vector because it is relatively safe and easy to use, has a broad host range and has a large capacity for dna inserts. Origins of truncated supplementary capsid proteins in. Us10017783b2 us14234,988 us201214234988a us10017783b2 us 10017783 b2 us10017783 b2 us 10017783b2 us 201214234988 a us201214234988 a us 201214234988a us 10017783 b2 us10017783 b2 us 10017783b2 authority us united states prior art keywords genome recombinant cell baculovirus genes prior art date 20110727 legal status the legal status is an assumption and is not. The indianmeal moth, plodia interpunctella lepidoptera. Use of whole genome sequence data to infer baculovirus.

Jul 24, 2007 baculovirus phylogeny based on complete genome sequence data. A number of genes lacking in acmnpv but present in other baculoviruses were. In this study, we have identified a pu sequence from the genome of a baculovirus which can strongly activate the activity of full or minimal promoters including the heat shock 70 promoter, the cmvm promoter, and minimal p35 promoters in insect cells. Improved generation of recombinant baculovirus genomes in. Phylogenetic analyses of these 30 genes yielded a tree with 4 major groups. The complete genome sequence of a single nucleocapsid nucleopolyhedrovirus recently isolated from chrysodeixis chalcites chchnpv was determined.

Comparison to other baculovirus genomes, hycugvhc1 revealed some differences in gene. Comparisons of 29 baculovirus genomes indicated that baculovirus phylogeny followed the classification of the hosts more closely than morphological traits that have previously been used for classification of this virus family. Arthropods, lepidoptera, hymenoptera, diptera, and decapoda serve as natural hosts. Replication of the baculovirus genome springerlink. The genome sequence of a polish isolate of agrotis segetum nucleopolyhedrovirus agsenpva was determined and analysed. The complete genome sequence of the perigonia luscaisolated group ii alphabaculovirus pelusnpv revealed that the virus is a new baculovirus species most closely related to clanis bilineata. In this study, genome sequences from 57 baculoviruses were analyzed to reevaluate the number and identity of core genes and to understand the distribution of the remaining coding sequences. Genome sequence of a baculovirus pathogenic for culex. However, this tool has not yet been adopted for use in the baculovirus insect cell system, which is an important recombinant protein production platform. Complete sequence and organization of antheraea pernyi. A betabaculovirus encoding a gp64 homolog bmc genomics. Immature larval forms of moth species are the most common hosts, but these viruses have also been. Kuzio j1, pearson mn, harwood sh, funk cj, evans jt, slavicek jm, rohrmann gf.