The nucleotide sequence from about 100 base-pairs downstream to about 600 base pairs upstream the... more The nucleotide sequence from about 100 base-pairs downstream to about 600 base pairs upstream the CloDF13 replication origin has been determined. A comparison of this sequence with the corresponding ColEl origin sequence reveals that:. The sequence at the origin of replication is conserved.. There are large differences in the nucleotide sequence downstream the replication origin, whereas there is a large homology in the region of about 410 base-pairs upstream the replication origin.. This conserved region might code for a largely homologous basic, arginine rich polypeptide of about 45 amino-acids, for both ColEl and CloDF13.. Although there are large differences in the primary structure of the region coding for the 100 nucleotide RNA, the secondary structure of this region seems to be conserved.
Previously, the gene for heat-stable enterotoxin II (STII) has been mapped by transposon Tn5 inse... more Previously, the gene for heat-stable enterotoxin II (STII) has been mapped by transposon Tn5 insertion mutagenesis in the chimeric R-Ent plasmid pCG86 (Mazaitis, A. J., R. Maas, and W. K. Maas, J. Bacteriol. 145:97-105, 1981). DNA segments containing this gene were cloned from the wild-type and STII-insertion-mutant plasmid. The position of the Tn5 insertion was determined, and a 530-base-pair-long segment of the wild-type plasmid corresponding to the Tn5 insertion site was sequenced. An open reading frame for the STII gene was identified and is characterized by typical promoter and ribosome binding site sequences. The deduced STII structural gene codes for a protein 71 amino acids long, including a typical signal peptide of 23 amino acids and a mature protein of 48 amino acids. The size and overall structure of STII are similar to those of STI, but the amino acid compositions of the two heat-stable enterotoxins are completely different.
XIth International Congress on Thrombosis and Haemostasis, 1987
The significance of the cleavage at arginine-275 of human t-PA has been the subject of debate. It... more The significance of the cleavage at arginine-275 of human t-PA has been the subject of debate. It has been reported, as expected for a member of the serine protease family, that the single chain form is a zymogen and that generation of catalytic activity is dependent upon cleavage at arginine-275. Other groups, in contrast, have found considerable enzyme activity associated with the one-chain form of t-PA. To clarify the functional significance of this proteolysis and circumvent cleavage of one-chain t-PA by itself or plasmin, site-directed mutagenesis was employed to change the codon of arginine-275 to specify a glutamic acid. The resulting plasmid was used to transfect CHO cells. The single chain mutant [Glu-275 t-PA] was expressed in CHO cells and the protein purified by conventional techniques. The mutant enzyme could be converted to the two-chain form by V8 protease, but not by plasmin. Glu-275 t-PA was 8 times less active in the cleavage of a tripeptide substrate and 20-50 tim...
Proceedings of the National Academy of Sciences, 1984
Plasmids have been constructed that direct the synthesis in Escherichia coli of heavy chains and/... more Plasmids have been constructed that direct the synthesis in Escherichia coli of heavy chains and/or light chains of an anti-carcinoembryonic antigen (CEA) antibody. Another plasmid was constructed for expression of a truncated form of heavy chain (Fd' fragment) in E. coli. Functional CEA-binding activity was obtained by in vitro reconstitution in E. coli extracts of heavy chain or Fd' fragment mixed with extracts containing light chain.
The nine base pairs immediately 5' of the initiation codon for the bovine growth hormone (BGH) st... more The nine base pairs immediately 5' of the initiation codon for the bovine growth hormone (BGH) structural gene have been mutagenized. The mutagenesis method employs the ligation of an ambiguously synthesized oligonucleotide duplex into a previously engineered gap in an expression plasmid for BGH. The mutation method, coupled with hybridization screening, is efficient at isolating 1 and 2 base pair changes within the targeted region. The E. coli cultures harboring the mutant plasmids were assayed for relative leveTs of BGH expression. The most notable result is the varied effect of substitution of G into the mRNA at various positions in this region.
The nucleotide sequence from about 100 base-pairs downstream to about 600 base pairs upstream the... more The nucleotide sequence from about 100 base-pairs downstream to about 600 base pairs upstream the CloDF13 replication origin has been determined. A comparison of this sequence with the corresponding ColEl origin sequence reveals that:. The sequence at the origin of replication is conserved.. There are large differences in the nucleotide sequence downstream the replication origin, whereas there is a large homology in the region of about 410 base-pairs upstream the replication origin.. This conserved region might code for a largely homologous basic, arginine rich polypeptide of about 45 amino-acids, for both ColEl and CloDF13.. Although there are large differences in the primary structure of the region coding for the 100 nucleotide RNA, the secondary structure of this region seems to be conserved.
This paper aims to investigate different factors determining strategic Decision making. This stud... more This paper aims to investigate different factors determining strategic Decision making. This study attempts to identify the role of RBV and RDT and its impact on strategic Decision making. It has been seen that different strategies has to be made to get competitive advantage keeping in mind external resources and while strategies differ when to get competitive advantage through internal resources such as Human resource and capital Resource. In this research heuristics and bounded rationality acted as moderator to strategic decision making as when decision makers (managers take decisions personal biases and heuristics also get involved.
In vitro recombination techniques were used to construct a series of new cloning vehicles with ge... more In vitro recombination techniques were used to construct a series of new cloning vehicles with genes of the tryptophan (trp) operon of Escherichia coli as selective marker. To construct these plasmids we have made a restriction cleavage map of the trp operon for the enzymes AosI, AvaI, BglI, BglII, HindIII, HpaI, PvuII, SalI, SstI and XhoI. The constructed plasmids pHP39, pEP392, pEP3921 and pEP3923 are derived from the amplifiable plasmid pBR345 and carry two or more genes of the trp operon, which are controlled by the trp regulatory elements. Plasmid pEP3921 (7.0 kb) carries intact trpE and trpA genes and contains single BglII and SstI sites in trpE, a single HindIII site located between trpE and trpA, and single EcoRI, SalI and XhoI sites located outside the trp genes. Plasmid pEP121 (12 kb) is similar to pEP3921, but has an extra selective marker conferring bacterial resistance to ampicillin. Plasmid pEP3923 (7.4 kb) comprises intact trpB and trpA genes and single BglII, HindIII, EcoRI, SalI and XhoI sites. Plasmids pHP39 (9.8 kb) and pEP392 (9.8 kb) carry an intact trp operon and have two and one EcoRI site, respectively. Plasmid pHP3 (18 kb) carries an intact trp operon and markers for tetracycline and ampicillin resistance.
cDNAs prepared using poly(A)mRNA from pituitaries and containing the coding sequences for bovine ... more cDNAs prepared using poly(A)mRNA from pituitaries and containing the coding sequences for bovine and porcine growth hormones (bGH and pGH) were cloned in bacteria. The primary structures of the peptide hormones derived from the nucleotide sequences of the respective cDNAs show approximately 90% homology. The cloned cDNAs were modified using synthetic DNA to construct expression vectors for efficient bacterial production of the mature animal growth hormones.
Tymosin fraction 5 is a partially purified bovine thymic preparation containing over 30 peptides ... more Tymosin fraction 5 is a partially purified bovine thymic preparation containing over 30 peptides with molecular weights ranging from 1000 to 15 000 (Hooper et al., 1975). Fraction 5 has been demonstrated to be an effective immunoptentiating agent and can act in lieu of the thymus ...
The objective of this section is to keep readers aware of significant inventions and trends in in... more The objective of this section is to keep readers aware of significant inventions and trends in industrial research as well as to highlight those areas of research that may lead to new biotechnological opportunities. Four major areas of biochemistry will be covered corresponding to enzymes, cells, bioproducts, and nucleic acids. The patent section will briefly cover each area in every issue of the journal. The literature section will focus on one area per issue. This section will identify patents and published patent applications from the international patent literature. The title, name(s) of the inventor(s), the patent number, the date of filing, the assignee, and a short description of the invention will be given. Copies of US patents can be obtained for 50r each from the Commissioner of Patents and Trademarks, Washington, DC 20231.
An abbreviated procedure has been developed for the purification of restriction endonucleases. Th... more An abbreviated procedure has been developed for the purification of restriction endonucleases. This procedure uses chromatography on phosphocellulose and hydroxylapatite and results in enzymes of sufficient purity to permit their use in the sequencing, molecular cloning, and physical mapping of DNA.
A method is described of cleaving double stranded DNA at any given point which comprises: (a) con... more A method is described of cleaving double stranded DNA at any given point which comprises: (a) converting the double stranded DNA to single-stranded DNA in a region surrounding the point; (b) hybridizing to the single-stranded region formed in step (a) a complementary primer length of single-stranded DNA, the 5' end of the primer lying opposite the nucleotide that is 5'
The nucleotide sequence from about 100 base-pairs downstream to about 600 base pairs upstream the... more The nucleotide sequence from about 100 base-pairs downstream to about 600 base pairs upstream the CloDF13 replication origin has been determined. A comparison of this sequence with the corresponding ColEl origin sequence reveals that:. The sequence at the origin of replication is conserved.. There are large differences in the nucleotide sequence downstream the replication origin, whereas there is a large homology in the region of about 410 base-pairs upstream the replication origin.. This conserved region might code for a largely homologous basic, arginine rich polypeptide of about 45 amino-acids, for both ColEl and CloDF13.. Although there are large differences in the primary structure of the region coding for the 100 nucleotide RNA, the secondary structure of this region seems to be conserved.
Previously, the gene for heat-stable enterotoxin II (STII) has been mapped by transposon Tn5 inse... more Previously, the gene for heat-stable enterotoxin II (STII) has been mapped by transposon Tn5 insertion mutagenesis in the chimeric R-Ent plasmid pCG86 (Mazaitis, A. J., R. Maas, and W. K. Maas, J. Bacteriol. 145:97-105, 1981). DNA segments containing this gene were cloned from the wild-type and STII-insertion-mutant plasmid. The position of the Tn5 insertion was determined, and a 530-base-pair-long segment of the wild-type plasmid corresponding to the Tn5 insertion site was sequenced. An open reading frame for the STII gene was identified and is characterized by typical promoter and ribosome binding site sequences. The deduced STII structural gene codes for a protein 71 amino acids long, including a typical signal peptide of 23 amino acids and a mature protein of 48 amino acids. The size and overall structure of STII are similar to those of STI, but the amino acid compositions of the two heat-stable enterotoxins are completely different.
XIth International Congress on Thrombosis and Haemostasis, 1987
The significance of the cleavage at arginine-275 of human t-PA has been the subject of debate. It... more The significance of the cleavage at arginine-275 of human t-PA has been the subject of debate. It has been reported, as expected for a member of the serine protease family, that the single chain form is a zymogen and that generation of catalytic activity is dependent upon cleavage at arginine-275. Other groups, in contrast, have found considerable enzyme activity associated with the one-chain form of t-PA. To clarify the functional significance of this proteolysis and circumvent cleavage of one-chain t-PA by itself or plasmin, site-directed mutagenesis was employed to change the codon of arginine-275 to specify a glutamic acid. The resulting plasmid was used to transfect CHO cells. The single chain mutant [Glu-275 t-PA] was expressed in CHO cells and the protein purified by conventional techniques. The mutant enzyme could be converted to the two-chain form by V8 protease, but not by plasmin. Glu-275 t-PA was 8 times less active in the cleavage of a tripeptide substrate and 20-50 tim...
Proceedings of the National Academy of Sciences, 1984
Plasmids have been constructed that direct the synthesis in Escherichia coli of heavy chains and/... more Plasmids have been constructed that direct the synthesis in Escherichia coli of heavy chains and/or light chains of an anti-carcinoembryonic antigen (CEA) antibody. Another plasmid was constructed for expression of a truncated form of heavy chain (Fd' fragment) in E. coli. Functional CEA-binding activity was obtained by in vitro reconstitution in E. coli extracts of heavy chain or Fd' fragment mixed with extracts containing light chain.
The nine base pairs immediately 5' of the initiation codon for the bovine growth hormone (BGH) st... more The nine base pairs immediately 5' of the initiation codon for the bovine growth hormone (BGH) structural gene have been mutagenized. The mutagenesis method employs the ligation of an ambiguously synthesized oligonucleotide duplex into a previously engineered gap in an expression plasmid for BGH. The mutation method, coupled with hybridization screening, is efficient at isolating 1 and 2 base pair changes within the targeted region. The E. coli cultures harboring the mutant plasmids were assayed for relative leveTs of BGH expression. The most notable result is the varied effect of substitution of G into the mRNA at various positions in this region.
The nucleotide sequence from about 100 base-pairs downstream to about 600 base pairs upstream the... more The nucleotide sequence from about 100 base-pairs downstream to about 600 base pairs upstream the CloDF13 replication origin has been determined. A comparison of this sequence with the corresponding ColEl origin sequence reveals that:. The sequence at the origin of replication is conserved.. There are large differences in the nucleotide sequence downstream the replication origin, whereas there is a large homology in the region of about 410 base-pairs upstream the replication origin.. This conserved region might code for a largely homologous basic, arginine rich polypeptide of about 45 amino-acids, for both ColEl and CloDF13.. Although there are large differences in the primary structure of the region coding for the 100 nucleotide RNA, the secondary structure of this region seems to be conserved.
This paper aims to investigate different factors determining strategic Decision making. This stud... more This paper aims to investigate different factors determining strategic Decision making. This study attempts to identify the role of RBV and RDT and its impact on strategic Decision making. It has been seen that different strategies has to be made to get competitive advantage keeping in mind external resources and while strategies differ when to get competitive advantage through internal resources such as Human resource and capital Resource. In this research heuristics and bounded rationality acted as moderator to strategic decision making as when decision makers (managers take decisions personal biases and heuristics also get involved.
In vitro recombination techniques were used to construct a series of new cloning vehicles with ge... more In vitro recombination techniques were used to construct a series of new cloning vehicles with genes of the tryptophan (trp) operon of Escherichia coli as selective marker. To construct these plasmids we have made a restriction cleavage map of the trp operon for the enzymes AosI, AvaI, BglI, BglII, HindIII, HpaI, PvuII, SalI, SstI and XhoI. The constructed plasmids pHP39, pEP392, pEP3921 and pEP3923 are derived from the amplifiable plasmid pBR345 and carry two or more genes of the trp operon, which are controlled by the trp regulatory elements. Plasmid pEP3921 (7.0 kb) carries intact trpE and trpA genes and contains single BglII and SstI sites in trpE, a single HindIII site located between trpE and trpA, and single EcoRI, SalI and XhoI sites located outside the trp genes. Plasmid pEP121 (12 kb) is similar to pEP3921, but has an extra selective marker conferring bacterial resistance to ampicillin. Plasmid pEP3923 (7.4 kb) comprises intact trpB and trpA genes and single BglII, HindIII, EcoRI, SalI and XhoI sites. Plasmids pHP39 (9.8 kb) and pEP392 (9.8 kb) carry an intact trp operon and have two and one EcoRI site, respectively. Plasmid pHP3 (18 kb) carries an intact trp operon and markers for tetracycline and ampicillin resistance.
cDNAs prepared using poly(A)mRNA from pituitaries and containing the coding sequences for bovine ... more cDNAs prepared using poly(A)mRNA from pituitaries and containing the coding sequences for bovine and porcine growth hormones (bGH and pGH) were cloned in bacteria. The primary structures of the peptide hormones derived from the nucleotide sequences of the respective cDNAs show approximately 90% homology. The cloned cDNAs were modified using synthetic DNA to construct expression vectors for efficient bacterial production of the mature animal growth hormones.
Tymosin fraction 5 is a partially purified bovine thymic preparation containing over 30 peptides ... more Tymosin fraction 5 is a partially purified bovine thymic preparation containing over 30 peptides with molecular weights ranging from 1000 to 15 000 (Hooper et al., 1975). Fraction 5 has been demonstrated to be an effective immunoptentiating agent and can act in lieu of the thymus ...
The objective of this section is to keep readers aware of significant inventions and trends in in... more The objective of this section is to keep readers aware of significant inventions and trends in industrial research as well as to highlight those areas of research that may lead to new biotechnological opportunities. Four major areas of biochemistry will be covered corresponding to enzymes, cells, bioproducts, and nucleic acids. The patent section will briefly cover each area in every issue of the journal. The literature section will focus on one area per issue. This section will identify patents and published patent applications from the international patent literature. The title, name(s) of the inventor(s), the patent number, the date of filing, the assignee, and a short description of the invention will be given. Copies of US patents can be obtained for 50r each from the Commissioner of Patents and Trademarks, Washington, DC 20231.
An abbreviated procedure has been developed for the purification of restriction endonucleases. Th... more An abbreviated procedure has been developed for the purification of restriction endonucleases. This procedure uses chromatography on phosphocellulose and hydroxylapatite and results in enzymes of sufficient purity to permit their use in the sequencing, molecular cloning, and physical mapping of DNA.
A method is described of cleaving double stranded DNA at any given point which comprises: (a) con... more A method is described of cleaving double stranded DNA at any given point which comprises: (a) converting the double stranded DNA to single-stranded DNA in a region surrounding the point; (b) hybridizing to the single-stranded region formed in step (a) a complementary primer length of single-stranded DNA, the 5' end of the primer lying opposite the nucleotide that is 5'
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Papers by H. Heyneker