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5
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3542995313
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thesis, University of Wisconsin-Madison
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D. J. Bowen, thesis, University of Wisconsin-Madison (1995).
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(1995)
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Bowen, D.J.1
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7
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0001652856
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V. J. Paul, S. Frautschy, W. Fenical, K. H. Nealson, J. Chem. Ecol. 7, 589 (1981).
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J. Chem. Ecol.
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Paul, V.J.1
Frautschy, S.2
Fenical, W.3
Nealson, K.H.4
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10
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0026466785
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S. Yamanaka, A. Hagiwara, Y. Nishimura, H. Tanabe, N. Ishibashi, Arch. Microbiol. 158, 387 (1992).
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(1992)
Arch. Microbiol.
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, pp. 387
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Yamanaka, S.1
Hagiwara, A.2
Nishimura, Y.3
Tanabe, H.4
Ishibashi, N.5
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14
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0025700045
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J. Van Rie, W. H. McGaughey, D. E. Johnson, B. D. Barnett, H. Van Mellaert, ibid. 247, 72 (1990).
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(1990)
Science
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Van Rie, J.1
McGaughey, W.H.2
Johnson, D.E.3
Barnett, B.D.4
Van Mellaert, H.5
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16
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3542992889
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note
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4, pH 6.9). Toxic fractions were concentrated, equilibrated with 10 mM tris-HCI (pH 8.6), and loaded onto a weak anion-exchange HPLC column (301VHP575) (Vydac) and eluted with a KCl gradient.
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17
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3542995908
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note
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2 cubes of artificial diet (Gypsy moth wheat germ diet; ICN Biomedical). For the range of other insects assayed (Table 1) either a mixture of the high molecular weight toxin complexes was added to the diet as for M. sexta or injectable activity was measured by injection of toxin in buffer directly into the insect hemocoel. For bioassay of the to gene disruptions, we used the same procedure except that instead of applying the same weight of purified Tc proteins, we added the same volume of different concentrations of culture broth to the diet cubes.
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19
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3543007383
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note
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Antisera were raised against the high molecular weight toxin fraction before further separation by HPLC. A rabbit polyclonal antiserum was raised against the native toxin, and a monoclonal antibody (C5F2) was derived from mice immunized with heat-denatured toxin.
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20
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3543046971
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+ (Stratagene) from size-fractionated DNA partially digested with Sau 3a and transformed into competent library efficiency XL2-Blue MRF' E. coli (Stratagene). The library was screened with both the monoclonal and the polyclonal antitoxin antibodies. Immunoreactive clones were restriction mapped and sequenced with Sequenase 2.0 (United States Biochemical).
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22
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3543009826
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personal communication
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D. Guiney, personal communication.
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Guiney, D.1
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23
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3543025359
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World Intellectual Property, Patent WO 97/17432 (1997)
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J. Ensign et al., World Intellectual Property, Patent WO 97/17432 (1997).
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Ensign, J.1
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0025977235
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J. C. Phelps, L. D. Lyerly, J. L. Johnson, T. D. Wilkins, Infect. Immun. 59, 150 (1991).
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(1991)
Infect. Immun.
, vol.59
, pp. 150
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Phelps, J.C.1
Lyerly, L.D.2
Johnson, J.L.3
Wilkins, T.D.4
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30
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0030894618
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C.-G. Yu, M. A. Mullins, G. W. Warren, M. G. Koziel, J. J. Estruch, Appl. Environ. Microbiol. 63, 532 (1997).
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(1997)
Appl. Environ. Microbiol.
, vol.63
, pp. 532
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Yu, C.-G.1
Mullins, M.A.2
Warren, G.W.3
Koziel, M.G.4
Estruch, J.J.5
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32
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3543029461
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For examination of gut pathology, we fed Tca toxin to neonate M. sexta for 3 hours and to first-instar larvae for 48 hours. Larvae were fixed in Bouin's fluid overnight at 4°C, dehydrated, and embedded in paraffin wax; 6-μm paraffin sections were stained with Weigert's hematoxylin followed by Cason's trichome stain; then they were examined by light microscopy.
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33
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3543000124
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Cells were spun down at 10,000g and then supernatants were concentrated in Millipore Centrifugal filtration units (Ultrafree, Biomax-100K) or diluted with growth medium.
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34
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3542997691
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note
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Abbreviations for the amino acid residues are as follows: A, Ala; C, Cys; D, Asp; E, Glu; F, Phe; G, Gly; H, His; I, Ile; K, Lys; L, Leu; M, Met; N, Asn; P, Pro; Q, Gln; R, Arg; S, Ser; T, Thr; V, Val; W, Trp; and Y, Tyr.
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3543003755
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note
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We thank all at Dow AgroSciences Biotechnology for their encouragement and support of this project. Supported by Hatch funds, The Applied Research and Technology Fund, and The Industrial and Economic Development Fund, all administered by the University of Wisconsin-Madison and by DowAgrosciences.
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