Biomedical subjects

G F Rohrmann

Publications and source records attributed to G F Rohrmann.

At least 73 records · Page 4Linked to original sources

PubMed1990

A capsid-associated protein of the multicapsid nuclear polyhedrosis virus of Orgyia pseudotsugata: genetic location, sequence, transcriptional mapping, and immunocytochemical characterization.

Two lambda gt11 clones containing overlapping DNA inserts encoding portions of a structural protein gene from Orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (OpMNPV) were identified by their immunoreactivity with polyclonal antisera produced against purified polyhedra-derived virus. Sequence analysis of a 3.6-kb region of the baculovirus genome (map units 69.1-71.6) from which the lambda gt11 inserts originated revealed an open reading frame of 1872 nt (624 amino acids) encoding a predicted protein of 70.6 kDa. Northern blot, primer extension, and 3' S1 analysis of this ORF indicated that an mRNA of approximately 2100 nt was transcribed from this gene. The mRNA appears to initiate from a late promoter/mRNA start site consensus sequence GTAAG and is expressed at late times postinfection. A gene fusion containing the C-terminal 368 amino acids of the gene was constructed using a bacterial trpE expression vector. Rabbit antiserum made against the purified fusion protein reacted with a protein of 87 kDa on Western blots of infected cell extracts at 24 hr p.i. and thereafter. The p87 protein was shown to be a component of both budded and polyhedra-derived virus and purified capsids. Immunofluorescence analysis indicated that p87 is expressed late in infection and concentrated in infected cell nuclei.

Amino Acid Sequence↗

PubMed1990

Characterization of the infection cycle of the Orgyia pseudotsugata multicapsid nuclear polyhedrosis virus in Lymantria dispar cells.

To characterize the infection cycle of the Orgyia pseudotsugata multicapsid nuclear polyhedrosis virus in Lymantria dispar cells, the time course of DNA synthesis and polyhedron production, and the onset and rate of budded virus production were investigated at three different m.o.i. (5, 10 and 100). In addition, the time course of expression of three proteins (gp64, p39 and polyhedrin) representative of three temporal classes of baculovirus genes was also analysed using Western blot analysis. DNA synthesis began at 12 to 18 h post-infection (p.i.). The rate of budded virus (BV) production reached maximal levels at 24 to 36 h p.i. and continued at high levels indicating that BV production was not turned off late in infection. Polyhedra were first observed at 48 h p.i. The m.o.i. appeared to influence the magnitude but not timing of early events in the viral infection cycle (gp64 expression and DNA synthesis) and also influenced the initial levels of BV production and the percentage of cells containing occlusion bodies. The m.o.i. had little influence on the final rates of BV production and the time of detection of p39 and polyhedrin on Western blots.

Animals↗

PubMed1990

The p6.5 gene region of a nuclear polyhedrosis virus of Orgyia pseudotsugata: DNA sequence and transcriptional analysis of four late genes.

The gene encoding the basic DNA-binding protein (p6.5) of the multicapsid nuclear polyhedrosis virus of Orgyia pseudotsugata (OpMNPV) was localized by Southern blot analysis using a cDNA probe containing the Autographa californica virus (AcMNPV) p6.9 gene. The OpMNPV p6.5 gene was mapped to the HindIII G fragment at map unit 67. Nucleotide sequence and transcriptional analysis of a 3.26 kb region encompassing this area revealed four open reading frames (ORFs 1 to 4) oriented in the same direction. ORF 1 demonstrated a seven codon overlap with ORF 2. Messenger RNAs initiated upstream of each of the four ORFs late in infection and were coterminal at a single site downstream of the fourth ORF. The conserved late gene promoter/mRNA start site sequence (ATAAG) was present upstream of all the ORFs, but did not appear to be the major site of mRNA initiation for the third ORF, as determined by primer extension analysis. The fourth ORF in this series encoded a predicted peptide of 51 amino acids (6.5K), which was 80% similar to the p6.9 basic DNA-binding protein of AcMNPV.

Amino Acid Sequence↗

PubMed1990

Baculovirus diversity and molecular biology.

With the identification and characterization of a number of structural and nonstructural protein genes, advances have been made in our understanding of baculovirus structure, regulation of gene expression, and replication. Since less than 30% of the AcMNPV genome has been sequenced and characterized, the continued identification and assignment of function to baculovirus genes is perhaps the most crucial of enterprises now facing baculovirologists and is critical to the development of our understanding of the baculovirus genome and its replication. The size and diversity of baculovirus genomes appears to be strongly influenced by mobile DNA from the insect host. Also, transposon-mediated mutations of baculoviruses provide examples of functional inactivation of viral genes (FP phenotype mutations) and transcriptional activation (TE-D insertion). Another role transposable elements may play is the introduction of insect promoters and enhancers to the baculovirus genome. Since early baculovirus genes are likely transcribed in a way similar to normal insect genes, transposons that insert strong constitutive promoters or cellular enhancers near early baculovirus genes may cause mutations that are subsequently selected for. If this does occur, baculovirus early gene promoters may exhibit a great deal of variability in sequence and may resemble host promoters. Given the overall similarity between the genomes of OpMNPV and AcMNPV and the apparent absence of a region, similar to the AcMNPV HindIII-K/EcoR1-S in OpMNPV, it is intriguing to speculate that this region which contains two ORFs and the hr5 enhancer, may have been inserted into the AcMNPV genome by transposition, possibly delivering several helpful genes (35k and 94k) and a powerful enhancer. The highly repeated enhancer may have been subsequently amplified by recombination. In such a model, the acquisition of general or species-specific enhancers might influence both virulence and host range. Acquisition of general enhancers could increase the level of early gene expression, thus accelerating the cellular infection cycle and making the virus more virulent. Similarly, the acquisition of species-specific enhancers might affect host range by accelerating the infection cycle, but only in a specific host or cell type. One might therefore postulate that diversity in baculoviruses may reflect not only different selection pressures but also the diversity of mobile DNA within host insect species. Although our understanding of baculovirus diversity and molecular biology is rapidly advancing, many of the fundamental characteristics that define the unique nature of baculoviruses remain poorly understood. One fundamental feature of baculoviruses is the production of the two virion phenotypes, PDV and BV.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1990

Mapping unprocessed epitopes using deletion mutagenesis of gene fusions.

To locate the antigenic determinant recognized by a monoclonal antibody directed against a baculovirus capsid protein, a series of overlapping deletions of a fusion protein were immunologically screened with the monoclonal antibody. The immunoreactive fusion protein was derived from a restriction fragment which contained a large portion of a baculovirus capsid protein open reading frame fused in-frame with a truncated trpE gene in a bacterial (pATH3) expression system. To map the epitope, nested sets of 5' and 3' deletion mutants were generated. Mutants were characterized by the DNA insert size or by the size of the expressed fusion protein. Selected N- and C-termini truncated fusion proteins were Western blotted and incubated with the monoclonal antibody to identify mutants which retained the epitope. Plasmid DNA from mutants which flank the 5' and 3' junction of the antigenic determinant were sequenced to determine the epitope junction. By screening forty 3' deletions and sixty-four 5' deletions, the antigenic determinant was localized to a region of seven amino acids.

Amino Acid Sequence↗

PubMed1989

A baculovirus polyhedral envelope-associated protein: genetic location, nucleotide sequence, and immunocytochemical characterization.

Using a polyclonal mouse antiserum produced against purified virions of the multicapsid nuclear polyhedrosis virus of Orgyia pseudotsugata (OpMNPV), two immunoreactive lambda gtII clones were identified which contained nonoverlapping insert DNAs which mapped to a single open reading frame (ORF) in the HindIII-M fragment. Analysis of nucleotide sequence data indicates that this ORF encodes a protein with a MW of 32.4 kDa. A trpE-p32 gene fusion containing the entire p32 ORF was constructed, and the fusion protein was purified and used to immunize rabbits. Western blot analysis and immunofluorescence studies using the anti-TrpE-p32 antiserum detected a polyhedra-derived virus (PDV)-associated protein of 32 kDa at 24 hr postinfection (hr p.i.). The protein was observed in the cytoplasm and nucleus at 24 hr p.i. and became concentrated in the cytoplasm late in infection. Western blot analysis and immunofluorescent microscopy of polyhedra solubilized under various conditions indicated that p32 is associated with the polyhedral envelope. The predicted amino acid sequence for p32 showed 58% amino acid identity with the predicted amino acid sequence for an ORF (ORF 3) in a similar region of the genome of the MNPV of Autographa californica (AcMNPV). The solubility properties of the p32 protein and reciprocal immunoblotting experiments indicate the OpMNPV p32 gene encodes a protein which is homologous to the polyhedral envelope-associated phosphoprotein of AcMNPV, pp34, recently reported by M.A. Whitt and J.S. Manning [(1988) Virology 163, 33-42].

Amino Acid Sequence↗

PubMed1989

Nucleotide sequence, transcriptional mapping, and temporal expression of the gene encoding p39, a major structural protein of the multicapsid nuclear polyhedrosis virus of Orgyia pseudotsugata.

The gene encoding the 39-kDa major structural protein (p39) of Orgyia pseudotsugata nuclear polyhedrosis virus (OpMNPV) was sequenced and transcriptionally mapped, and its expression was examined at various times postinfection. By Northern hybridization, primer extension, and S1 nuclease analysis, we identified p39 mRNAs of approximately 2600 nt. By primer extension analysis, we identified two major sets of transcripts which initiated around -48 and -96 nt upstream of the translation start codon. The transcription start sites were located within the conserved baculovirus late gene consensus sequence, ATAAG, which is duplicated in the p39 5' flanking region. In OpMNPV-infected Lymantria dispar cells, the p39 mRNAs were expressed abundantly at 24 and 36 hr p.i. but were present in lower quantities at 48 hr p.i. The p39 gene contained an open reading frame of 1053 nt which encodes a predicted protein of 351 amino acids with an estimated molecular weight of 39.5 kDa. Three repeats of the amino acid sequence Ala-Pro-Ala-Ala-Pro were identified at the C-terminus of the predicted p39 protein.

Amino Acid Sequence↗

PubMed1989

Location, sequence, transcriptional mapping, and temporal expression of the gp64 envelope glycoprotein gene of the Orgyia pseudotsugata multicapsid nuclear polyhedrosis virus.

The gene encoding gp64, the envelope glycoprotein of the budded virus (BV) of Orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (OpMNPV), was mapped to the HindIII-E fragment of of the viral genome and expression of the gp64 gene was examined at various times postinfection. To locate the gp64 gene, a cross-reacting monoclonal antibody (AcV5) (A. W. Hohmann and P. Faulkner, 1983, Virology, 125, 432-444) directed against the gp64 protein of the Autographa californica multicapsid nuclear polyhedrosis virus (AcMNPV) was used to screen a lambda gt11 expression library of OpMNPV and insert DNAs from immunopositive recombinants were used for Southern hybridization mapping. The gp64 gene was sequenced and transcription of the gp64 gene was examined by Northern blot, S1 nuclease, and primer extension analysis. Two sets of gp64 transcripts were detected during infection: a single early transcript which initiated at -43 nt and four late transcripts which initiated at -152, -167, -174, and -175 nt relative to the start of the gp64 open reading frame. Comparison of the gp64 early transcription initiation site with several other early baculovirus genes revealed a four-nucleotide consensus sequence (CAGT) which is conserved at the early transcription initiation sites of the IE-1 and 39K genes. The four late gp64 transcripts initiated at two of the four upstream ATAAG motifs. All gp64 mRNAs appear to be coterminal at the 3' end. Analysis of the nucleotide sequence of the gp64 gene revealed that the late gp64 mRNAs are bicistronic, consisting of a three amino acid minicistron located 70 nt upstream of the 509 amino acid gp64 open reading frame. Early transcripts do not contain the minicistron. The 1527-nt gp64 open reading frame encodes a predicted protein of 509 amino acids with a molecular weight of 58 kDa. The predicted gp64 protein contains seven potential N-linked glycosylation sites and hydrophobic N- and C-termini characteristic of signal and membrane anchor sequences found on envelope glycoproteins. By western blot analyses and indirect immunofluorescence microscopy, we show that the gp64 protein is present at early times (6 hr) postinfection and accumulates in the infected cell, moving to the periphery at later times postinfection. Western blot comparisons of the temporal expression of the gp64 protein with the p39 capsid protein revealed that these two virion structural protein genes differ significantly in the timing of their initial expression. The upstream regulatory regions, open reading frames, and predicted proteins from the OpMNPV and AcMNPV gp64 genes were compared.(ABSTRACT TRUNCATED AT 400 WORDS)

Amino Acid Sequence↗

PubMed1989

Characterization of the genetic organization of the HindIII M region of the multicapsid nuclear polyhedrosis virus of Orgyia pseudotsugata reveals major differences among baculoviruses.

A region including the 4 kb HindIII M fragment of the multicapsid nuclear polyhedrosis virus (MNPV) of Orgyia pseudotsugata (OpMNPV) genome was sequenced, transcriptionally mapped, and compared to the homologous region in the MNPV of Autographa californica (AcMNPV). Five open reading frames (ORFs) oriented in the same direction were identified and were found to be expressed at late times post-infection. The mRNAs from the five ORFs were found to coterminate at a single site downstream of ORF 5. The conserved late gene promoter/mRNA start site sequence (AGTAAG) was present upstream of all the ORFs, but did not appear to be the major site of mRNA initiation for two of the ORFs as determined by primer extension analysis. These data indicated that use of this signal for transcriptional initiation may vary between different ORFs. The predicted amino acid sequences for the five ORFs of AcMNPV and OpMNPV were compared and amino acid homologies of 26 to 72% were observed. The comparison revealed a number of major differences in the genomes of the two viruses. A putative transposable element of 634 nucleotides was found to be inserted into the previously reported AcMNPV ORF 1 sequence. In addition, it was found that a region corresponding to the 4 kb HindIII K/EcoRI S/hr5 region of AcMNPV was not present in the OpMNPV genome.

Amino Acid Sequence↗

PubMed1988

The genome of the multicapsid baculovirus of Orgyia pseudotsugata: restriction map and analysis of two sets of GC-rich repeated sequences.

Five cosmids containing inserts that comprise the complete genome of the multicapsid nuclear polyhedrosis virus of Orgyia pseudotsugata were mapped with four restriction enzymes (Bg/II, ClaI, SstI, XhoI). From these cosmid maps, composite maps of the complete genome were constructed for each restriction enzyme. A region containing repeats of the sequence GGC downstream of the polyhedrin gene was used to probe the genome. It cross-hybridized with a region which, upon sequence analysis, was found to be a highly repetitive GC-rich region of nearly 500 nucleotides. The two GC-rich regions appeared to be evolutionarily unrelated.

Animals↗

PubMed1988

p39, a major baculovirus structural protein: immunocytochemical characterization and genetic location.

A series of monoclonal antibodies was produced against proteins from polyhedra-derived virions of the multicapsid nuclear polyhedrosis virus of Orgyia pseudotsugata (OpMNPV). Two of these antibodies (214 and 236) reacted with a protein of 39 kDa on Western blots of electrophoretically separated OpMNPV viron proteins derived from both budded and polyhedra-derived virions. This protein appears to be a major component of both BV and PDV. One of the p39 antibodies was used to characterize p39 synthesis in infected Lymantria dispar cells by using Western blots and immunofluorescent staining. The p39 protein was detected by immunofluorescence microscopy at 24 hr postinfection. By 48 hr, p39 was detected primarily in cell nuclei with little or no detectable staining of the cytoplasm. The two MAbs were used to identify three immunoreactive clones from a lambda gt11 expression library of OpMNPV DNA. By hybridization of insert DNA from three lambda gt11 clones to blots of restriction digests of OpMNPV genomic DNA, the location of the 39-kDa gene was mapped on the OpMNPV genome. Using the lambda gt11 insert DNAs and the monoclonal antibodies, the p39 genes and proteins of OpMNPV and Autographa californica NPV (AcMNPV) were shown to be closely related in size, DNA sequence, and antigenicity. One of the p39 monoclonal antibodies cross-reacted with a host cell protein associated with the condensed chromosomes present during mitosis.

Antibodies, Monoclonal↗

PubMed1987

Characterization of baculovirus p10 synthesis using monoclonal antibodies.

A series of monoclonal antibodies were produced against virion proteins of the multicapsid nuclear polyhedrosis virus of Orgyia pseudotsugata (OpMNPV). Four of these antibodies reacted with a protein of 14 kd on Western blots of electrophoretically separated OpMNPV virion proteins. These antibodies were used to identify immunoreactive clones from a lambda gt11 expression library of OpMNPV DNA. By hybridization of insert DNA from the lambda gt11 clones to blots of digests of OpMNPV genomic DNA, and by sequencing the ends of the lambda gt11 inserts, these clones were shown to contain a portion of the p10 gene. The regions containing epitopes recognized by the four monoclonal antibodies were located using fusion proteins made from selected portions of the p10 reading frame in a trpE vector. One of the p10 antibodies was used to characterize p10 synthesis in infected Lymantria dispar cells by using Western blots and immunofluorescent staining. The p10 protein was detected with immunofluorescent microscopy at 14 hr postinfection and by 20 hr it formed intensely staining cytoplasmic structures. On Western blots of infected cells, two forms of p10 (of about 14 and 15 kd) were observed. One of the p10 monoclonal antibodies showed a strong cross-reaction with cytoskeletal structures in uninfected insect cells and rat fibroblasts.

Animals↗

PubMed1987

Comparison of the p26 gene region of two baculoviruses.

A 1.1-kb region of DNA containing the p26 gene of the multicapsid nuclear polyhedrosis virus of Orgyia pseudotsugata (OpMNPV) was sequenced, transcriptionally mapped, and compared to the same region in the MNPV of Autographa californica (AcMNPV). The mRNA start site of the p26 gene occurs about 22 nucleotides downstream from an A/T-rich putative promoter sequence that is highly conserved between AcMNPV and OpMNPV. The p26 mRNA is transcribed through the p26 gene and coterminates with the p10 gene resulting in a mRNA containing copies of both genes. The reading frames of the OpMNPV and AcMNPV p26 genes showed 47% amino acid sequence homology which is clustered in six regions with over 65% amino acid homology. There was a distinct bias toward incorporation of G/C-rich codons in the OpMNPV p26 gene. No DNA homology was observed between the region upstream of the p26 gene in AcMNPV and OpMNPV. In AcMNPV, this region contains the homologous repeated (hr) sequence hr5. Hybridization of a plasmid containing an AcMNPV-repeated sequence (hr5) to Southern blots of the OpMNPV genome indicated that this repeated sequence is lacking in OpMNPV.

Amino Acid Sequence↗

PubMed1986

Nucleotide sequencing and transcriptional mapping of the Orgyia pseudotsugata multicapsid nuclear polyhedrosis virus p10 gene.

A 32P-labeled cloned DNA fragment (AcMNPV HindIII-Q) containing one of the repeated sequences and a portion of the p10 gene from Autographa californica multicapsid nuclear polyhedrosis virus (AcMNPV) was used to probe Southern blots containing restriction endonuclease digests of Orgyia pseudotsugata muticapsid nuclear polyhedrosis virus (OpMNPV) DNA. A single 3.6-kb fragment, OpMNPV HindIII-Q, was hybridized. The OpMNPV HindIII-Q fragment was cloned into pUC-18, mapped with restriction endonucleases, and reprobed with the AcMNPV HindIII-Q fragment. A small region of ca. 700 bp, near the left end of the cloned fragment, was cross-hybridized. DNA sequencing in this region revealed an open reading frame of 279 bp which shares detectable homology with the p10 gene of AcMNPV. The sequences downstream from the p10 gene in both viruses also contain long open reading frames which share homology. Northern blot analysis of RNA from OpMNPV infected O. leucostigma cells was used to define the temporal and spatial organization of transcripts from this region. S1 analysis of both termini of the major p10 mRNA indicates nontranslated regions of 52-53 bases at the 5' end and 175 bases at the 3' end. The 5'-mRNA start site was located within a 12-nucleotide sequence which is conserved in all late hyperexpressed baculovirus genes.

Animals↗

PubMed1986

Polyhedrin structure.

Polyhedrin has evolved two highly specialized functions. Firstly, it forms a protective crystal around the virus and secondly it resists solubilization except under strongly alkaline conditions similar to those found in the insect midgut. Both of these properties allow the virus to remain viable for many years outside the insect host. Although polyhedrin and granulin can vary by about 50% in amino acid sequence, many of their structural features are highly conserved, reflecting the similar function and biochemical properties of these proteins. By comparing the sequences, domains within the gene which evolve rapidly have been identified and molecular phylogenies have been proposed. Information on predicted secondary structure has also been obtained and some insight into the possible role of codon usage in baculovirus function has also been gained from the sequence information. In addition to the conserved structural properties of the polyhedrin protein, there is a conserved regulatory process which results in the synthesis of massive amounts of polyhedrin. This process is probably governed by a virus-specific RNA polymerase. A potential regulatory signal shared by all these genes has been identified upstream from the polyhedrin gene. A number of additional granulin and lepidopteran polyhedrin sequences will certainly be forthcoming because of the ease with which these genes are identified by cross-hybridization with available related probes. However, of special interest will be sequences from dipteran and hymenopteran polyhedrins which will add greatly to our understanding of the constraints governing polyhedrin structure and diversity. Another logical step in their study will be to examine polyhedrin quaternary structure utilizing X-ray crystallography. Additional areas of future emphasis will probably focus on the regulation of polyhedrin synthesis. Elucidation of the regulatory signals governing transcription of these genes are of prime interest as are complementary studies on the characterization of the RNA polymerase which transcribes these genes.

Amino Acid Sequence↗

PubMed1985

Cloning and sequencing of the granulin gene from the Trichoplusia ni granulosis virus.

A restriction fragment containing the granulin gene from the Trichoplusia ni granulosis virus was located in a blot of viral genomic DNA using a cloned polyhedrin gene as a probe. This fragment was cloned, mapped, subcloned, and the sequence of the coding region and 5' and 3' flanking regions was determined. Although granulin is very similar in size to nuclear polyhedrosis virus polyhedrins, the N-terminal region of granulin demonstrated a high degree of variability with the first 60 amino acids only 28% homologous to the Autographa californica nuclear polyhedrosis virus (AcMNPV) polyhedrin sequence. Between amino acid 60 and the carboxyterminus at amino acid 248, the sequence was very similar (64%) to polyhedrin sequences. Overall the nucleotide and amino acid sequences were 58 and 53%, respectively, related to those of AcMNPV. No introns were evident in the gene.

Amino Acid Sequence↗

PubMed1985

The nucleotide sequence of the Pieris brassicae granulosis virus granulin gene.

Two overlapping restriction fragments containing the Pieris brassicae granulosis virus (GV) granulin gene were cloned into plasmids. The regions containing the coding region and the 5' and 3' flanking regions were subcloned into M13 and sequenced. The nucleotide sequence data were compared to those for the granulin gene from the Trichoplusia ni GV and the polyhedrin gene from the Autographa californica nuclear polyhedrosis virus (AcMNPV). The amino acid sequences derived from these DNA sequences indicated that the two GV proteins are more closely related to each other (77% amino acid homology) than either is to the AcMNPV (about 53% amino acid homology for either GV). The N-terminal region shows the greatest degree of variation between these proteins. Highly conserved amino acid sequences were identified between the two GVs and were also found between NPVs. Certain of these conserved regions are shared between GVs and NPVs while others are not.

Amino Acid Sequence↗

PubMed1985

Delta endotoxin of Bacillus thuringiensis subsp. israelensis.

From Bacillus thuringiensis subsp. israelensis, a proteinase-resistant protein was purified which exhibited toxicity to larval mosquitoes and cultured mosquito cells, lysed erythrocytes, and was lethal to mice. To extract the protein, a sporulating culture of B. thuringiensis subsp. israelensis was treated with alkali, neutralized, and incubated with trypsin and proteinase K. It was then purified by gel filtration and DEAE column chromatography. Up to 240 micrograms of toxic protein was purified from 1 g (wet weight) of culture pellet. Two closely related forms of toxic protein were obtained: the 25a and 25b proteins. The two forms comigrated near 25,000 daltons in a sodium dodecyl sulfate-polyacrylamide gel, were serologically related, and showed similar partial protease digestion profiles, but were distinguishable by DEAE chromatography and nondenaturing polyacrylamide gel electrophoresis. Protein sequencing data indicated the 25b protein lacked the two amino acids at the amino terminus of the 25a protein. A Western blot enzyme-linked immunosorbent assay of alkali-solubilized proteins that were not treated with proteases suggested the toxic 25a and 25b proteins were proteolytically derived from a larger molecule of about 28,000 daltons. Alkali-solubilized proteins from an acrystalliferous strain of B. thuringiensis subsp. israelensis and from B. thuringiensis subsp. kurstaki failed to cross-react with antibodies to the 25a protein.

Amino Acid Sequence↗