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  • 1
    Electronic Resource
    Electronic Resource
    [s.l.] : Nature Publishing Company
    Nature biotechnology 15 (1997), S. 887-890 
    ISSN: 1546-1696
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: [Auszug] Antifreeze proteins (AFP) inhibit ice growth by surface adsorption that results in a depression of the freezing point below the melting point. The maximum level of this thermal hysteresis shown by the four structurally unrelated fish AFP is approximately 1.5°C. In contrast, hemolymph and crude ...
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  • 2
    Electronic Resource
    Electronic Resource
    [s.l.] : Nature Publishing Company
    Nature biotechnology 8 (1990), S. 453-457 
    ISSN: 1546-1696
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: [Auszug] We have expressed two antifreeze protein genes from the Atlantic wolffish, Anarhichas lupus, in Drosophila melanogaster by placing them under the divergent transcriptional control of the host yolk poly-peptide (1 and 2) gene promoters. Both genes were joined to the central promoter region by fusion ...
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  • 3
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] Insect antifreeze proteins (AFP) are considerably more active at inhibiting ice crystal growth than AFP from fish or plants. Several insect AFPs, also known as thermal hysteresis proteins, have been cloned and expressed. Their maximum activity is 3–4 times that of fish AFPs and they ...
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  • 4
    Electronic Resource
    Electronic Resource
    [s.l.] : Macmillan Magazines Ltd.
    Nature 388 (1997), S. 727-728 
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] We have purified a thermal hysteresis (antifreeze) protein, with up to 100 times the specific activity of fish antifreeze proteins, from the common yellow mealworm beetle, Tenebrio molitor. It is a threonine- and cysteine-rich protein, of relative molecular mass 8,400, composed largely of ...
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  • 5
    Electronic Resource
    Electronic Resource
    [s.l.] : Nature Publishing Group
    Nature 361 (1993), S. 470-472 
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] TPI was purified to homogeneity from fourth instar larval homogenates by successive steps of ion exchange, ultrafiltration,gel filtration chromatography, and fast protein liquid chromatography (FPLC) Mono-Q (Pharmacia). Its identity was confirmed by enzyme assays, amino-terminal sequencing, and ...
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Biochemical genetics 23 (1985), S. 363-378 
    ISSN: 1573-4927
    Keywords: Drosophila melanogaster ; Minute mutations ; yolk polypeptides
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Minutes have been considered for some time to be mutant at the sites of synthesis of some components of the protein synthetic apparatus. To study the hypothetical relationship between Minutes and suboptimal translation, a group of abundant proteins, the yolk polypeptides, was assayed in outcrossed females bearing M(3)w, M(3)h y , or M(1)n mutations. Recently emerged Minute females contained a lower amount of yolk polypeptides, in both ovarian and nonovarian tissues, than their non-Minute sisters. This low level correlated with the lower abundance of cytoplasmic RNA in Minutes compared to control females. By 1 week of age, both M(3)w and their non-Minute sibs contained the same amount of yolk polypeptides and the corresponding mRNA. The double heterozygote, ap 4/+;M(3)w/+, did not differ in yolk polypeptide content from control flies. M(3)w females demonstrated reduced fecundity during the period of low yolk polypeptide content but gradually increased egg deposition as yolk polypeptide levels rose. These results suggest that the low protein levels are due to the slower maturation of M(3)w, and not to less efficient translation machinery.
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Biochemical genetics 35 (1997), S. 119-138 
    ISSN: 1573-4927
    Keywords: insecticide resistance ; mosquitoes ; esterase ; cDNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract A malathion-resistant strain of Culex tarsalis has a malathion carboxylesterase which rapidly hydrolyzes the insecticide. This is in contrast to organophosphate-resistant strains of C. quinquefasciatus and C. pipiens, which have elevated levels of general B esterases due to amplification of the corresponding genes, producing increased amounts of enzyme which appear to protect the insects by sequestering the insecticide. The contribution to resistance of the homologous esterase B3 (Estβ3) gene (estβ3) in C. tarsalis was investigated by cloning and characterizing sequences from resistant and susceptible strains. estβ3 is similar to estβ1, both structurally and in sequence. The first intron of estβ3,however, has a region of extensive repeats which may be responsible for the inefficient processing of the transcript. Southern blots indicate that the gene is single copy in both strains, and northern blots show that it is not greatly overexpressed in the resistant insects. estβ3 cDNAs from resistant and susceptible strains have 98% amino acid identity. It appears that, in contrast to other studies, estβ3 does not play a significant role in insecticide resistance in our strains of C. tarsalis, and the molecular responses of pest insects to organophosphates may be more diverse than has been suggested.
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Biochemical genetics 19 (1981), S. 47-60 
    ISSN: 1573-4927
    Keywords: Drosophila melanogaster ; aminopeptidases ; leucine aminopeptidases
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Two leucine aminopeptidases from Drosophila melanogaster larvae have been partially purified. The LAP A and D enzymes have similar biochemical characteristics including molecular weights of ≅280,000 daltons, Michaelis-Menten constants of ≅0.05 mM, associations with metal cofactors, and specificities toward natural and chromogenic substrates. They differ in their pH optima and spatial distributions. If the closely linked genes that code for these enzymes have resulted from a tandem gene duplication event, it is suggested that there has been subsequent evolutionary divergence. This would provide Drosophila larvae with two related, but functionally distinct enzymes.
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Biochemical genetics 32 (1994), S. 9-24 
    ISSN: 1573-4927
    Keywords: insecticide resistance ; malathion carboxylesterase ; Lucilia cuprina
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Resistance to the organophosphorus insecticide malathion in genetically related strains of the Australian sheep blowflyLucilia curprina was examined. Separate lines of blowflies were established by homozygosis of the fourth chromosome of the parental RM strain. Both the RM and the derived resistant (der-R) strains are approximately 100 times more resistant to malathion than the related susceptible der-S strain, resistance being correlated with a 45- to 50-fold increase in a malathion carboxylesterase (MCE) activity. MCE has a pH optimum ranging between 6.6 and 8.0 and is strongly inhibited by the carboxylesterase inhibitors triphenyl phosphate, paraoxon, and diiospropylfluorophosphate. Subcellular fractionation revealed that MCE was localized predominantly to the cytosol and mitochondria in both resistant and susceptible blowflies. A single MCE was purified to homogeneity from RM blowflies. It has a pI of 5.5, is a monomer of 60.5 kDa, and hydrolyzes malathion with aV max of 755 nmol/min/mg protein and aK m of 11.0 µM. L. cuprina have thus evolved a remarkable MCE which is faster and more efficient at hydrolyzing a specific insecticide than any other insect esterase yet described.
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  • 10
    ISSN: 1573-9368
    Keywords: proprotein ; secretion ; haemolymph ; thermal hysteresis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have usedDrosophila melanogaster as a model system for the transgenic expression of cystine-rich Type II antifreeze protein (AFP) from sea raven. This protein was synthesized and secreted into fly haemolymph where it migrated as a larger species (16 kDa) than the mature form of the protein (14 kDa) as judged by immunoblotting.Drosophila-produced Type II AFP demonstrated antifreeze activity both in terms of thermal hysteresis (0.13 °C) and inhibition of ice recrystallization. Recombinant AFP was purified and N-terminal sequencing revealed a 17 aa extension that began at the predicted signal peptide cleavage point. The expression of all three AFP types in transgenicDrosophila has now been achieved. We conclude that the globular Type II and Type III AFPs are better choices for antifreeze transfer to other organisms than is the more widely used linear Type I AFP.
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