Gerrit Veeneman - Academia.edu (original) (raw)
Papers by Gerrit Veeneman
Journal of Biological Chemistry, 1987
Journal of Food Protection, 1989
An agglutination test by using latex beads (0.8 μm diameter) coated with IgG from the antibodies ... more An agglutination test by using latex beads (0.8 μm diameter) coated with IgG from the antibodies against extra-cellular polysaccharide of Penicillium digitatum has been developed. As low as 5 to 10 ng/ml of the purified extra-cellular polysaccharide of the same species can be detected by this preparation. Analysis of culture filtrates from 25 different molds showed that the positive reactivity was obtained only with the species of genera Penicillium and Aspergillus. The application of this test was confirmed in testing the samples of spices and nuts. Further, the reliability could be enhanced by including specific blocker in the assay, the synthetic epitopes consisting of four β (1–5)-linked D-galactofuranosyl residues.
Advances in Experimental Medicine and Biology, 1984
Synthetic oligodeoxyribonucleotides, DNA ligase and DNA polymerase were used to construct double-... more Synthetic oligodeoxyribonucleotides, DNA ligase and DNA polymerase were used to construct double-stranded DNA fragments homologous to the first 25, 27 or 30 b.p. of the 30 b.p. origin region of bacteriophage phi X174 (nucleotides 4299-4328 of the phi X174 DNA sequence). The double-stranded DNA fragments were cloned into the kanamycin resistance gene of pACYC177 (AmpR, KmR). Transformants were picked up by antibiotic selection and filter-hybridization using one of the oligodeoxyribonucleotides as a probe. Approximate lengths of the inserts were determined by restriction enzyme analysis. Exact length and orientation of each insert was determined by DNA sequencing. Plasmid DNA with an insert homologous to the first 25 b.p. of the phi X174 origin is not nicked by the gene A protein. However, plasmid DNA containing the 27 b.p. fragment in either orientation is nicked by the gene A protein, as well as plasmid DNAs containing the first 28 b.p. or the complete 30 b.p. conserved origin region of the isometric phages.
ChemInform, 2010
ABSTRACT ChemInform is a weekly Abstracting Service, delivering concise information at a glance t... more ABSTRACT ChemInform is a weekly Abstracting Service, delivering concise information at a glance that was extracted from about 100 leading journals. To access a ChemInform Abstract of an article which was published elsewhere, please select a “Full Text” option. The original article is trackable via the “References” option.
Recueil des Travaux Chimiques des Pays-Bas, 2010
ABSTRACT The partially protected and crystalline 2′-O-levulinoyl-3′5′-O-(tetraisopropyldisiloxane... more ABSTRACT The partially protected and crystalline 2′-O-levulinoyl-3′5′-O-(tetraisopropyldisiloxane-1,3-diyl)guanosine derivative (4) serves as a key intermediate for the synthesis of 2′-O-acetal-N2-acylriboguanosines 1c. Acylation of 4 with diphenylacetic anhydride gave, after hydrazinolysis of the levulinoyl group, the N2-acyl derivative 6 in high yield. Acylation of 4 with benzoyl or biphenyl-4-carbonyl chloride afforded the corresponding N2-acyl derivatives in lower yields. Acetalization of derivatives 6 with 4-methoxy-5,6-dihydro-2H-pyran, followed by the removal of the 1,1,3,3-tetraisopropyldisiloxane-1,3-diyl (TIPS) group, gave the corresponding 2′-O-acetal-N2-acylriboguanosines 1c.
European Journal of Biochemistry, 1980
The deoxyoctanucleotide (5'-3')d(A-A-G-G-A-G-G-T), which is complementary to the 3' end of 16-S R... more The deoxyoctanucleotide (5'-3')d(A-A-G-G-A-G-G-T), which is complementary to the 3' end of 16-S RNA, inhibits the formation of the complex between the 30-S subunit and MS2 RNA described in the preceding paper. If the complex is preformed, the octanucleotide cannot prevent entry of the complex into the ribosome cycle upon supplementation with the components for protein synthesis. The subunit. MS2-RNA complex is unable to bind the octanucleotide. It is concluded that in the subunit. phage-RNA initiation precursor the 1 6 3 terminus is base-paired with a complementary MS2 RNA sequence. Edeine, aurintricarboxylic acid and antibodies against ribosomal protein S1 prevent the association of phage RNA with 30-S subunits. These compounds do not, however, inhibit the binding of (5'-3')d(A-A-G-G-A-G-G-T) to 30-S subunits. It is concluded that the formation of the complex between MS2 RNA and 30-S subunits does not depend solely on the Shine and Dalgarno base-pairing reaction.
Tetrahedron Letters, 1985
... A VERSATILE PHOSPHORYLATING AGENT * CTJ Wreesmann, A. Fidder, GH Veeneman, GAvan der Marel an... more ... A VERSATILE PHOSPHORYLATING AGENT * CTJ Wreesmann, A. Fidder, GH Veeneman, GAvan der Marel and dH van Boom Gorl aeus ... 4 (1 mmol), which was previously dried by coevapora tion with pyridine, in dioxan was added to a stirred solution of 3 (1.1 nimol) . ...
Tetrahedron Letters, 1981
ABSTRACT Morpholino phosphorobis-3-nitro-1,2,4-triazolidate () has been applied to the phosphoryl... more ABSTRACT Morpholino phosphorobis-3-nitro-1,2,4-triazolidate () has been applied to the phosphorylation of partially protected (5′-OH free) DNA intermediates (). Using this procedure, the following types of compounds were synthesized: pBp(Bp)nB; AppBp(Bp)nB and ppBp(Bp)nB.
Tetrahedron, 1992
The disaccharide a-o-GlcNAcp-(1-5)-~-KW-2-OCH,CH,CH@I~ containing a spacer B-linlred to KDO is pq... more The disaccharide a-o-GlcNAcp-(1-5)-~-KW-2-OCH,CH,CH@I~ containing a spacer B-linlred to KDO is pqanzd via iodonium ion-assisted glycosylation of a suitable KDO acceptor with ethyl 2-azido-2~xy-3,4~-~-~yl-l-~~~-~ glucopyranoside. The key KDO building unit is synthesized in 7 steps starting with 23:5.6-di-O-isoproW1idene-~m~i~I.
Nucleic Acids Research, 1984
Synthetic oligonucleotides, DNA ligase and DNA polymerase were used to construct double-stranded ... more Synthetic oligonucleotides, DNA ligase and DNA polymerase were used to construct double-stranded DNA fragments homologous to the first 25, 27 or 30 b.p. of the origin of replication of bacteriophage 0X174 (nucleotides 4299-4328 of the 0X174 DNA sequence). The double-stranded DNA fragments were cloned into the unique SmaI or R HindIII restriction sites in the kanamycin-resistance gene of pACYCI77 (Amp KmR). Recombinant plasmids were picked up by colony hybridization. DNA sequencing showed that not only recombinant plasmids with the expected insert were formed, but also recombinant plasmids with a shorter insert. Recombinant plasmids with an insert homologous to the first 24, 25, 26, 27, 28 or all 30 b.p. of the 0X174 origin region were thus obtained. Supercoiled plasmids containing a sequence homologous to the first 27, 28 or 30 b.p. of the 0X174 origin region are nicked by the 0X174 gene A protein. However, the other supercoiled plasmids are not nicked by the 0X174 gene A protein. These results show that the first 27 b.p. of the 0X174 origin region are sufficient as well as required for the initiation step in 0X174 RF DNA replication, i.e. the cleavage by gene A protein.
Nucleic Acids Research, 1986
The DNA from various human tumors and tumor cell lines was screened for the presence of mutated r... more The DNA from various human tumors and tumor cell lines was screened for the presence of mutated ras oncogenes with synthetic oligonucleotide probes, as well as with the NIH/3T3 cell transfection assay. Among the various mutations found we discovered two novel Ki-ras mutations in codon 12: gly to ala and gly to ser. A gastric carcinoma was found to possess a single mutated Ki-ras allele (gly-12 to ser), as well as a 30-50 fold amplified normal allele. This implies that two activating steps must have occurred in this malignancy.
Nucleic Acids Research, 1984
The activation of ras genes in naturally occurring tumors has, thus far, been found to be due to ... more The activation of ras genes in naturally occurring tumors has, thus far, been found to be due to mutations in codon 12 or 61 resulting in single amino acid substitutions. We have used highly labeled synthetic oligonucleotides to detect mutations in these codons and to determine the exact position of the mutation. Using this approach we have found three different mutations in codon 61 of the N-ras gene of various human tumor cell lines. In the fibrosarcoma line HT1080 the first nucleotide of the codon is mutated; in the promyelocytic line HL60 the second and in the rhabdomyosarcoma line RD301 the third nucleotide. For RD301 this implies that the normal glutamine residue at position 61 is replaced by histidine. In addition to the mutated N-ras gene the three cell lines have a normal N-ras gene which is indicative of the dominant character of the mutations.
Nucleic Acids Research, 1980
Nucleotide sequences at the 5'-termini of the alfalfa mosaic virus genomic RNAs and the intercist... more Nucleotide sequences at the 5'-termini of the alfalfa mosaic virus genomic RNAs and the intercistronic junction in RNA 3 were deduced and compared to identify possible common recognition signals for replicating enzymes in the corresponsing minusstranded viral RNAs. Homology between the 51-terminal sequences is less than 11 nucleotides and no complementarity with the homologous sequence occurring at the 3'-end of the viral RNAs was observed. Homology between the 51-terminus and intercistronic region in RNA 3 is compatible with the synthesis of subgenomic RNA 4 by internal initiation of transcription on the RNA 3 minusstrands. The sequence around the intercistronic junction can be folded into a very stable secondary structure.
Nucleic Acids Research, 1984
Phosphorylation of N-acyl-5'-0-DMTr-d-nucleosides, or similarly protected DNA-dimers having a fre... more Phosphorylation of N-acyl-5'-0-DMTr-d-nucleosides, or similarly protected DNA-dimers having a free 3'-OH group, with 2-chlorophenyl-0,0-bis(1-benzotriazolyl)phosphate affords reactive 3'-phosphotriester derivatives. The latter intermediates can be used, without further purification, for the synthesis of DNA-fragments on the controlled pore glass/long chain alkylamine support. Further, 2-cyano-1,1-dimethylethoxy dichlorophosphine proved to be very suitable for the preparation of 5'-phosphorylated DNA-fragments on the same type of solid support.
Nature, 1980
ABSTRACT Gene A protein, the initiator protein of bacteriophage PhiX174 DNA replication, cleaves ... more ABSTRACT Gene A protein, the initiator protein of bacteriophage PhiX174 DNA replication, cleaves synthetic single-stranded oligodeoxyribonucleotides at the same site as the corresponding sequence at the PhiX origin. The results identify the recognition sequence within the decamer CAACTTGATA which is cleaved next to the G residue. Further requirements for cleavage of double-stranded DNA by the gene A protein are supercoiling and an A + T-rich domain adjacent to the recognition sequence.
Molecular Immunology, 1988
Aspergillus and Penicillium species produce extracellular polysaccharides which are immunological... more Aspergillus and Penicillium species produce extracellular polysaccharides which are immunologically active. Methyl b-D-galactofuranoside interferes with the reaction between the polysaccharide antigens and the antibodies raised in rabbits. Of the different interlinked dimers of p-D-galactofuranosides (l-2; l-3; l-+5; l-*6) the (l-5) interlinked p-D-galactofuranoside gave the highest inhibition. An increasing inhibitory effect of di-, tri-, tetra-, penta-, hexa-, and heptamers of (1+5) interlinked B-D-galactofuranosides was observed. It was noticed that the penta-, hexa-and heptamer of (l-5) interlinked b-D-galactofuranosides were able to link antibodies raised against the extracellular polysaccharides produced by Penicillium species. The tetramer molecule was able to neutralize the binding of antibodies, which are naturally present in human sera, to the polysaccharides produced by Penicillium and Aspergillus species.
Journal of Molecular Biology, 1983
We have studied the activation of genes for VSGs (variant surface glycoproteins) in Trypanosoma b... more We have studied the activation of genes for VSGs (variant surface glycoproteins) in Trypanosoma brucei (strain 427
Journal of Molecular Biology, 1981
FEBS Letters, 1982
The bacteriophage 9X174 gene A encodes two proteins: gene A protein and A* protein. Purified A* p... more The bacteriophage 9X174 gene A encodes two proteins: gene A protein and A* protein. Purified A* protein acts as a single-stranded, DNA-specific endonuclease which remains covalently attached to the 5'-end of the cleavage site. Incubation of A* protein with the synthetic heptamer CAACTTG or with oligonucleotides which yield this heptamer after cleavage with the A* protein yields oligonucleotides with the sequences CAACTTGAG, CAACTTGAGG and CAACTTGAGGA. This indicates that A* protein carries an oligonucleotide with the sequence-AG,-AGG or-AGGA. The oligonucleotide can be transferred to the 3'-end of the heptamer CAACTTG. This suggests that A* protein reacts with a specific DNA sequence in the infected cell. Bacteriophage 4x174 A * protein DNA-protein complex Synthetic oligonucleotide DNA-ligating activity
Brain Research, 1986
The possibility has been mentioned that a change in the structure is responsible for the deviant ... more The possibility has been mentioned that a change in the structure is responsible for the deviant behavioral activity of y-endorphin in extracts of postmortem brain and pituitary gland samples of schizophrenic patients. This paper describes the investigation of this possibility by means of: (1) amino acid composition analysis of a-and 7-endorphin isolated from a pituitary gland of a schizophrenic patient; and (2) nucleotide sequence analysis of the 7-endorphin coding region of pro-opiomelanocortin (POMC) mRNA from two other pituitary glands, using the primer extension method. Both methods require no more than a single pituitary to obtain reliable results, a-and ~,-endorphin were isolated from an acid extract by gel filtration and two subsequent HPLC steps. In addition, the 7-endorphin region of fl-endorphin was analyzed by enzymatic cleavage of fl-endorphin and isolation of the resulting fragment. Single-stranded y-endorphin cDNA was synthesized by reverse transcriptase using total cellular pituitary RNA and a 5' 32p-labeled oligodeoxyribonucleotide primer (20-mer) hybridizing close to the y-endorphin coding region of POMC mRNA. Single-stranded cDNA was digested with restriction enzyme HaeIII which generated a 148 nucleotides long radioactive cDNA fragment containing the 7-endorphin cDNA sequence. The sequence of the 148 nucleotides fragment was determined. Neither the amino acid composition analysis nor the amino acid sequence derived from the cDNA nucleotide sequence revealed differences between schizophrenics and controls. Thus, no evidence was found for changes in the amino acid sequence of pituitary y-endorphin in these analyses, which include 3 cases of schizophrenia. Our structural data exclude the possibility of a mutation in the y-endorphin coding region of both POMC alleles. Also no evidence was obtained for the heterozygotic situation, i.e. the situation in which only one of the two alleles contains a mutation.
Journal of Biological Chemistry, 1987
Journal of Food Protection, 1989
An agglutination test by using latex beads (0.8 μm diameter) coated with IgG from the antibodies ... more An agglutination test by using latex beads (0.8 μm diameter) coated with IgG from the antibodies against extra-cellular polysaccharide of Penicillium digitatum has been developed. As low as 5 to 10 ng/ml of the purified extra-cellular polysaccharide of the same species can be detected by this preparation. Analysis of culture filtrates from 25 different molds showed that the positive reactivity was obtained only with the species of genera Penicillium and Aspergillus. The application of this test was confirmed in testing the samples of spices and nuts. Further, the reliability could be enhanced by including specific blocker in the assay, the synthetic epitopes consisting of four β (1–5)-linked D-galactofuranosyl residues.
Advances in Experimental Medicine and Biology, 1984
Synthetic oligodeoxyribonucleotides, DNA ligase and DNA polymerase were used to construct double-... more Synthetic oligodeoxyribonucleotides, DNA ligase and DNA polymerase were used to construct double-stranded DNA fragments homologous to the first 25, 27 or 30 b.p. of the 30 b.p. origin region of bacteriophage phi X174 (nucleotides 4299-4328 of the phi X174 DNA sequence). The double-stranded DNA fragments were cloned into the kanamycin resistance gene of pACYC177 (AmpR, KmR). Transformants were picked up by antibiotic selection and filter-hybridization using one of the oligodeoxyribonucleotides as a probe. Approximate lengths of the inserts were determined by restriction enzyme analysis. Exact length and orientation of each insert was determined by DNA sequencing. Plasmid DNA with an insert homologous to the first 25 b.p. of the phi X174 origin is not nicked by the gene A protein. However, plasmid DNA containing the 27 b.p. fragment in either orientation is nicked by the gene A protein, as well as plasmid DNAs containing the first 28 b.p. or the complete 30 b.p. conserved origin region of the isometric phages.
ChemInform, 2010
ABSTRACT ChemInform is a weekly Abstracting Service, delivering concise information at a glance t... more ABSTRACT ChemInform is a weekly Abstracting Service, delivering concise information at a glance that was extracted from about 100 leading journals. To access a ChemInform Abstract of an article which was published elsewhere, please select a “Full Text” option. The original article is trackable via the “References” option.
Recueil des Travaux Chimiques des Pays-Bas, 2010
ABSTRACT The partially protected and crystalline 2′-O-levulinoyl-3′5′-O-(tetraisopropyldisiloxane... more ABSTRACT The partially protected and crystalline 2′-O-levulinoyl-3′5′-O-(tetraisopropyldisiloxane-1,3-diyl)guanosine derivative (4) serves as a key intermediate for the synthesis of 2′-O-acetal-N2-acylriboguanosines 1c. Acylation of 4 with diphenylacetic anhydride gave, after hydrazinolysis of the levulinoyl group, the N2-acyl derivative 6 in high yield. Acylation of 4 with benzoyl or biphenyl-4-carbonyl chloride afforded the corresponding N2-acyl derivatives in lower yields. Acetalization of derivatives 6 with 4-methoxy-5,6-dihydro-2H-pyran, followed by the removal of the 1,1,3,3-tetraisopropyldisiloxane-1,3-diyl (TIPS) group, gave the corresponding 2′-O-acetal-N2-acylriboguanosines 1c.
European Journal of Biochemistry, 1980
The deoxyoctanucleotide (5'-3')d(A-A-G-G-A-G-G-T), which is complementary to the 3' end of 16-S R... more The deoxyoctanucleotide (5'-3')d(A-A-G-G-A-G-G-T), which is complementary to the 3' end of 16-S RNA, inhibits the formation of the complex between the 30-S subunit and MS2 RNA described in the preceding paper. If the complex is preformed, the octanucleotide cannot prevent entry of the complex into the ribosome cycle upon supplementation with the components for protein synthesis. The subunit. MS2-RNA complex is unable to bind the octanucleotide. It is concluded that in the subunit. phage-RNA initiation precursor the 1 6 3 terminus is base-paired with a complementary MS2 RNA sequence. Edeine, aurintricarboxylic acid and antibodies against ribosomal protein S1 prevent the association of phage RNA with 30-S subunits. These compounds do not, however, inhibit the binding of (5'-3')d(A-A-G-G-A-G-G-T) to 30-S subunits. It is concluded that the formation of the complex between MS2 RNA and 30-S subunits does not depend solely on the Shine and Dalgarno base-pairing reaction.
Tetrahedron Letters, 1985
... A VERSATILE PHOSPHORYLATING AGENT * CTJ Wreesmann, A. Fidder, GH Veeneman, GAvan der Marel an... more ... A VERSATILE PHOSPHORYLATING AGENT * CTJ Wreesmann, A. Fidder, GH Veeneman, GAvan der Marel and dH van Boom Gorl aeus ... 4 (1 mmol), which was previously dried by coevapora tion with pyridine, in dioxan was added to a stirred solution of 3 (1.1 nimol) . ...
Tetrahedron Letters, 1981
ABSTRACT Morpholino phosphorobis-3-nitro-1,2,4-triazolidate () has been applied to the phosphoryl... more ABSTRACT Morpholino phosphorobis-3-nitro-1,2,4-triazolidate () has been applied to the phosphorylation of partially protected (5′-OH free) DNA intermediates (). Using this procedure, the following types of compounds were synthesized: pBp(Bp)nB; AppBp(Bp)nB and ppBp(Bp)nB.
Tetrahedron, 1992
The disaccharide a-o-GlcNAcp-(1-5)-~-KW-2-OCH,CH,CH@I~ containing a spacer B-linlred to KDO is pq... more The disaccharide a-o-GlcNAcp-(1-5)-~-KW-2-OCH,CH,CH@I~ containing a spacer B-linlred to KDO is pqanzd via iodonium ion-assisted glycosylation of a suitable KDO acceptor with ethyl 2-azido-2~xy-3,4~-~-~yl-l-~~~-~ glucopyranoside. The key KDO building unit is synthesized in 7 steps starting with 23:5.6-di-O-isoproW1idene-~m~i~I.
Nucleic Acids Research, 1984
Synthetic oligonucleotides, DNA ligase and DNA polymerase were used to construct double-stranded ... more Synthetic oligonucleotides, DNA ligase and DNA polymerase were used to construct double-stranded DNA fragments homologous to the first 25, 27 or 30 b.p. of the origin of replication of bacteriophage 0X174 (nucleotides 4299-4328 of the 0X174 DNA sequence). The double-stranded DNA fragments were cloned into the unique SmaI or R HindIII restriction sites in the kanamycin-resistance gene of pACYCI77 (Amp KmR). Recombinant plasmids were picked up by colony hybridization. DNA sequencing showed that not only recombinant plasmids with the expected insert were formed, but also recombinant plasmids with a shorter insert. Recombinant plasmids with an insert homologous to the first 24, 25, 26, 27, 28 or all 30 b.p. of the 0X174 origin region were thus obtained. Supercoiled plasmids containing a sequence homologous to the first 27, 28 or 30 b.p. of the 0X174 origin region are nicked by the 0X174 gene A protein. However, the other supercoiled plasmids are not nicked by the 0X174 gene A protein. These results show that the first 27 b.p. of the 0X174 origin region are sufficient as well as required for the initiation step in 0X174 RF DNA replication, i.e. the cleavage by gene A protein.
Nucleic Acids Research, 1986
The DNA from various human tumors and tumor cell lines was screened for the presence of mutated r... more The DNA from various human tumors and tumor cell lines was screened for the presence of mutated ras oncogenes with synthetic oligonucleotide probes, as well as with the NIH/3T3 cell transfection assay. Among the various mutations found we discovered two novel Ki-ras mutations in codon 12: gly to ala and gly to ser. A gastric carcinoma was found to possess a single mutated Ki-ras allele (gly-12 to ser), as well as a 30-50 fold amplified normal allele. This implies that two activating steps must have occurred in this malignancy.
Nucleic Acids Research, 1984
The activation of ras genes in naturally occurring tumors has, thus far, been found to be due to ... more The activation of ras genes in naturally occurring tumors has, thus far, been found to be due to mutations in codon 12 or 61 resulting in single amino acid substitutions. We have used highly labeled synthetic oligonucleotides to detect mutations in these codons and to determine the exact position of the mutation. Using this approach we have found three different mutations in codon 61 of the N-ras gene of various human tumor cell lines. In the fibrosarcoma line HT1080 the first nucleotide of the codon is mutated; in the promyelocytic line HL60 the second and in the rhabdomyosarcoma line RD301 the third nucleotide. For RD301 this implies that the normal glutamine residue at position 61 is replaced by histidine. In addition to the mutated N-ras gene the three cell lines have a normal N-ras gene which is indicative of the dominant character of the mutations.
Nucleic Acids Research, 1980
Nucleotide sequences at the 5'-termini of the alfalfa mosaic virus genomic RNAs and the intercist... more Nucleotide sequences at the 5'-termini of the alfalfa mosaic virus genomic RNAs and the intercistronic junction in RNA 3 were deduced and compared to identify possible common recognition signals for replicating enzymes in the corresponsing minusstranded viral RNAs. Homology between the 51-terminal sequences is less than 11 nucleotides and no complementarity with the homologous sequence occurring at the 3'-end of the viral RNAs was observed. Homology between the 51-terminus and intercistronic region in RNA 3 is compatible with the synthesis of subgenomic RNA 4 by internal initiation of transcription on the RNA 3 minusstrands. The sequence around the intercistronic junction can be folded into a very stable secondary structure.
Nucleic Acids Research, 1984
Phosphorylation of N-acyl-5'-0-DMTr-d-nucleosides, or similarly protected DNA-dimers having a fre... more Phosphorylation of N-acyl-5'-0-DMTr-d-nucleosides, or similarly protected DNA-dimers having a free 3'-OH group, with 2-chlorophenyl-0,0-bis(1-benzotriazolyl)phosphate affords reactive 3'-phosphotriester derivatives. The latter intermediates can be used, without further purification, for the synthesis of DNA-fragments on the controlled pore glass/long chain alkylamine support. Further, 2-cyano-1,1-dimethylethoxy dichlorophosphine proved to be very suitable for the preparation of 5'-phosphorylated DNA-fragments on the same type of solid support.
Nature, 1980
ABSTRACT Gene A protein, the initiator protein of bacteriophage PhiX174 DNA replication, cleaves ... more ABSTRACT Gene A protein, the initiator protein of bacteriophage PhiX174 DNA replication, cleaves synthetic single-stranded oligodeoxyribonucleotides at the same site as the corresponding sequence at the PhiX origin. The results identify the recognition sequence within the decamer CAACTTGATA which is cleaved next to the G residue. Further requirements for cleavage of double-stranded DNA by the gene A protein are supercoiling and an A + T-rich domain adjacent to the recognition sequence.
Molecular Immunology, 1988
Aspergillus and Penicillium species produce extracellular polysaccharides which are immunological... more Aspergillus and Penicillium species produce extracellular polysaccharides which are immunologically active. Methyl b-D-galactofuranoside interferes with the reaction between the polysaccharide antigens and the antibodies raised in rabbits. Of the different interlinked dimers of p-D-galactofuranosides (l-2; l-3; l-+5; l-*6) the (l-5) interlinked p-D-galactofuranoside gave the highest inhibition. An increasing inhibitory effect of di-, tri-, tetra-, penta-, hexa-, and heptamers of (1+5) interlinked B-D-galactofuranosides was observed. It was noticed that the penta-, hexa-and heptamer of (l-5) interlinked b-D-galactofuranosides were able to link antibodies raised against the extracellular polysaccharides produced by Penicillium species. The tetramer molecule was able to neutralize the binding of antibodies, which are naturally present in human sera, to the polysaccharides produced by Penicillium and Aspergillus species.
Journal of Molecular Biology, 1983
We have studied the activation of genes for VSGs (variant surface glycoproteins) in Trypanosoma b... more We have studied the activation of genes for VSGs (variant surface glycoproteins) in Trypanosoma brucei (strain 427
Journal of Molecular Biology, 1981
FEBS Letters, 1982
The bacteriophage 9X174 gene A encodes two proteins: gene A protein and A* protein. Purified A* p... more The bacteriophage 9X174 gene A encodes two proteins: gene A protein and A* protein. Purified A* protein acts as a single-stranded, DNA-specific endonuclease which remains covalently attached to the 5'-end of the cleavage site. Incubation of A* protein with the synthetic heptamer CAACTTG or with oligonucleotides which yield this heptamer after cleavage with the A* protein yields oligonucleotides with the sequences CAACTTGAG, CAACTTGAGG and CAACTTGAGGA. This indicates that A* protein carries an oligonucleotide with the sequence-AG,-AGG or-AGGA. The oligonucleotide can be transferred to the 3'-end of the heptamer CAACTTG. This suggests that A* protein reacts with a specific DNA sequence in the infected cell. Bacteriophage 4x174 A * protein DNA-protein complex Synthetic oligonucleotide DNA-ligating activity
Brain Research, 1986
The possibility has been mentioned that a change in the structure is responsible for the deviant ... more The possibility has been mentioned that a change in the structure is responsible for the deviant behavioral activity of y-endorphin in extracts of postmortem brain and pituitary gland samples of schizophrenic patients. This paper describes the investigation of this possibility by means of: (1) amino acid composition analysis of a-and 7-endorphin isolated from a pituitary gland of a schizophrenic patient; and (2) nucleotide sequence analysis of the 7-endorphin coding region of pro-opiomelanocortin (POMC) mRNA from two other pituitary glands, using the primer extension method. Both methods require no more than a single pituitary to obtain reliable results, a-and ~,-endorphin were isolated from an acid extract by gel filtration and two subsequent HPLC steps. In addition, the 7-endorphin region of fl-endorphin was analyzed by enzymatic cleavage of fl-endorphin and isolation of the resulting fragment. Single-stranded y-endorphin cDNA was synthesized by reverse transcriptase using total cellular pituitary RNA and a 5' 32p-labeled oligodeoxyribonucleotide primer (20-mer) hybridizing close to the y-endorphin coding region of POMC mRNA. Single-stranded cDNA was digested with restriction enzyme HaeIII which generated a 148 nucleotides long radioactive cDNA fragment containing the 7-endorphin cDNA sequence. The sequence of the 148 nucleotides fragment was determined. Neither the amino acid composition analysis nor the amino acid sequence derived from the cDNA nucleotide sequence revealed differences between schizophrenics and controls. Thus, no evidence was found for changes in the amino acid sequence of pituitary y-endorphin in these analyses, which include 3 cases of schizophrenia. Our structural data exclude the possibility of a mutation in the y-endorphin coding region of both POMC alleles. Also no evidence was obtained for the heterozygotic situation, i.e. the situation in which only one of the two alleles contains a mutation.