Improved apparatus for vertical gel electrophoresis

Roberts, Richard M.; Jones, Joshua S.

doi:10.1016/0003-2697(72)90469-1

Cited by 17 publications

(3 citation statements)

References 5 publications

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“…Esterases were examined with the technique of polyacrylamide slab gel electrophoresis described by Hubby and Lewontin (1966), with the following• modifications: the age of the flies varied from 1 day to 70 days; flies were ground in 15 or 20 !J-l of homogenizing buffer (0.1 l\1 tris-borate with EDTA, pH 8.9), but the amount of sucrose in the buffer was changed from 5% to 10% halfway through the experimentation; from 5 to 10 !J-l of uncentrifuged homogenate was layered in the gel pocket. During the course of experimentation, a new apparatus was designed and employed to speed gTeatly the process of preparing and loading the gels (Roberts 1971;Roberts and Jones 1972) and was manufactured by Aardvark Instruments, r~ombard, Illinois. After electrophoresis, gels were immersed in 0.5 M boric acid solution at room temperature for 10-20 min, before incubating for 1-4 hr at 21°-29° C in 100 ml of 0.1 111 phosphate buffer, pH 6.5, containing 25 mg a-naphthyl acetate and 25 mg ~-naphthyl acetate, and 50 mg Fast Blue BB (from Dajac I1aboratories, Philadelphia).…”

Section: Electrophoresismentioning

confidence: 99%

Frequency Distribution and Linkage Disequilibrium of Active and Null Esterase Isozymes in Natural Populations of Drosophila montana

Roberts¹,

Baker²

1973

The American Naturalist

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Section: Electrophoresismentioning

confidence: 99%

Frequency Distribution and Linkage Disequilibrium of Active and Null Esterase Isozymes in Natural Populations of Drosophila montana

Roberts¹,

Baker²

1973

The American Naturalist

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“…In particular, we found that digestion of dilute preparations of intact virions (*«75 nm diameter) in the presence of Na+ or Li+ ions results in their conversion to intermediate subviral particles («* 65 nm diameter) which demonstrate enhanced infectivity, selectively altered polypeptide composition and latent transcriptase activity. Digestion(w/v) in bromphenol blue dye, and suitable aliquots were loaded into the sample wells in a slab gel which had been cast in an apparatus described by R oberts and Jones [8]. For routine analyses the separating gel composition was 7.5% (w/v) acrylamide-bisacrylamide (19:1) (Cyanogum-41, purchased from Isolab, Inc., Akron, Ohio), 0.1% (w/v) SDS, 6 m urea, 0.10 m Tris-Cl (pH 8.3), 0.02 m EDTA, 0.16% (w/v) ammonium persulfate and 0.025% (v/v) TEMED.…”

mentioning

confidence: 99%

Reovirus Transcriptase Activation in vitro: Further Studies on the Facilitation Phenomenon

Borsa¹,

Dg²,

Sargent³

et al. 1974

Intervirology

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Activation of the transcriptase in purified reovirus by chymotrypsin requires facilitating conditions in the digestion mix. In the presence of facilitation the virions are uncoated to cores. Without facilitation, uncoating is blocked at an intermediate stage, with transcriptase latent. Two independent types of facilitation have been discovered. The first type is mediated by K⁺, Rb⁺ or Cs⁺ ions. A second type, described here, is mediated by suitably high concentrations of virus in the digestion mix and requires collisions between intermediate subviral particles for its occurrence. The virus-concentration-mediated process is reversibly inhibited by high levels of active chymotrypsin.

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“…Electromorphs were distinguished and aphids then assigned to clone groups according to their combination of electromorphs at the two loci. Details of the electrophoresis were as described previously (Baker, 1977; Hubby & Lewontin, 1966; Roberts & Jones, 1974) but with one improvement: 0.01 M sodium glycinate, pH 8.8, was used as gel, electrode and homogenisation buffer in place of tris maleate EDTA. Use of this glycine buffer significantly increased the activity of the resistance associated esterase, RAE.…”

Section: A Na Iysismentioning

confidence: 99%

Electrophoretic studies on populations of Myzjus persicae in Scotland from March to July, 1976

Baker

1978

Annals of Applied Biology

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S U M M A R YSpecimens of Myzus persicae have been collected from field populations in Scotland and scored electrophoretically for their phenotypes at three polymorphic loci. These phenotypes were almost invariably present in individuals in one of a small number of favoured combinations, suggesting that these populations may be oligogeneous, that is, composed of large numbers of copies of a few favoured combinations of alleles.Temporal and spatial sampling at sites in East and West Scotland in 1976 has shown that different allele combination types were present in the two areas. Populations in E. Scotland contained very few specimens belonging to the organophosphorus (OP) resistant clone types (using esterase activity to infer resistance); instead there were large numbers of an OP susceptible type which has not been detected in England, but which came to predominate on cabbage and potato host plants at every site sampled during the period of rapid population increase. In contrast, populations in W. Scotland were characterised by substantial numbers of specimens belonging to OP resistant clone types and by an almost complete absence of this susceptible type. Two other distinguishable OP susceptible types were found to be common to both areas. The W. Scotland populations closely resemble N. England populations while those in E. Scotland appear to be unique to that area. Specimens of OP resistant types were only detected in E. Scotland after the beginning of June and in such relative proportions as to strongly suggest that they were migrants from W. Scotland or N. England. J . P . B A K E R M A T E R I A L S A N D M E T H O D S SamplingPopulations of M. persicae were sampled at a number of sites in Scotland and resampled at 3weekly intervals for as long as possible.Overwintering populations were sampled on 22 March at four sites that had not been treated with insecticide, in the Edinburgh-E. Lothian region ( Table 1). These were at Musselburgh (W l ) , Cockburnspath (W2), Haddington (W3) and Bo'ness (W4). At three of these sites the crops were lifted soon after sampling and the fields ploughed, but at the fourth (site W l), the host plants (cabbage) were left for a hrther 3 months and five successive aphid samples were collected at 3-weekly intervals from the site ( Table 2). 4 8 9 21 June 17 9 20 16 45Summer populations of M. persicae were first sampled in June 1976 on potato at five sites in the Edinburgh-E. Lothian area, four near Haddington and one, Site 5, at Edinburgh airport; at two sites at Invergowrie in the Dundee area, and at three sites 2 4 miles east of Prestwick in Ayrshire. Three weeks later a second sample was taken from these Sites, and at six of them it was possible to collect a third sample a further 3 wk later (Tables 3-5). At the time of first sampling none of these populations had been treated with insecticides, but two had been treated with OP's by the time the second sample was collected.Sampling methods were as previously described (Baker, 1977), namely, only one aphid per plant and plants ch...

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Improved apparatus for vertical gel electrophoresis

Cited by 17 publications

References 5 publications

Frequency Distribution and Linkage Disequilibrium of Active and Null Esterase Isozymes in Natural Populations of Drosophila montana

Frequency Distribution and Linkage Disequilibrium of Active and Null Esterase Isozymes in Natural Populations of Drosophila montana

Reovirus Transcriptase Activation in vitro: Further Studies on the Facilitation Phenomenon

Electrophoretic studies on populations of Myzjus persicae in Scotland from March to July, 1976

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