Quantitation of chloramphenicol acetyl transferase in transgenic tobacco plants by ELISA and correlation with gene copy number

A monoclonal antibody to chloramphenicol acetyl transferase (CAT) was used in an indirect competitive enzyme immunoassay (ELISA) for the quantitation of CAT in leaf extracts of eighteen transgenic tobacco plants containing the CAT gene fused to the cauliflower mosaic virus 35S promoter. The ELISA could be used to quantify CAT when present in extracts at 20 ng/ml. Enzymatic activity and electrophoretic mobility of CAT in these extracts was not different from CAT from Escherichia coli. Concentrations of CAT in these transgenic plants ranged from 79 to 732 ng CAT/mg protein. The average coefficient of variation among three replicate samples was 15%. All plants were sampled on two separate occasions. The CAT concentrations often varied between the two sampling dates. We determined the CAT gene copy number and the number of independently segregating loci in each plant by Southern blot analysis and progeny testing. We found no significant differences in CAT expression among all ten plants with a single CAT gene. We also found a significant correlation between CAT gene copy number and the level of CAT expressed in each plant, although plants with one gene copy sometimes had more CAT than plants with more than one gene copy. In this population, therefore, gene copy number contributed more to the variation in CAT expression than did position effects.     

Fecundity and egg fertility in the adult Colorado beetle (Leptinotarsa decemlineata) surviving larval infection by the fungus Beauveria bassiana

Secondary effects of the entomopathogenic hyphomycete Beauveria bassiana (Bals.) Vuill. were investigated on adults of Leptinotarsa decemlineata Say (Coleoptera:Chrysomelidae) treated as newly moulted 4th-instar larvae. Fecundity and egg hatching were monitored in surviving females mated with surviving males. When reared at 22°C, adult survivors showed a lessening of their reproductive potential during their whole life. According to the fungal inoculum dosage, the reductions of the total number of eggs laid per female and the mean number of eggs per egg mass ranged from 20% to 56% and from 18% to 46%, respectively. In contrast, at 25°C, the fecundity of survivors was not affected by the fungal infection. This variability of the secondary effects of B. bassiana according to temperature conditions might be related to the lower infection level of treated larvae at 25°C. Moreover, at both temperatures, eggs laid by females surviving fungal infection as larvae were as fertile as eggs laid by control insects. It is concluded that secondary effects of B. bassiana on the fecundity of the Colorado Potato Beetle are temperature-dependent and that they could become negligible at high temperatures under field conditions.

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Résumé Les effets secondaires de l'hyphomycète entomopathogène, Beauveria bassiana (Bals.) Vuill. sur la fécondité et la fertilité des insectes survivant à l'infection fongique ont été étudiés chez les adultes du doryphore Leptinotarsa decemlineata Say (Coleoptera:Chrysomelidae) issus de larves contaminées en début de 4ème stade. Les insectes maintenus à 22°C ont présenté une réduction de leur potentiel reproductif. Ainsi, suivant la dose d'inoculum fongique, le nombre total d'oeufs pondus par femelle et le nombre moyen d'oeufs par opplaque ont baissé de 20% à 56% et de 18% à 46%, respectivement. En revanche, à 25°C la fécondité des adultes survivants n'a pas été affectée par la maladie. Cette variabilité en fonction des conditions thermiques peut être liée à la diminution de l'effet-dose du champignon sur les larves à 25°C par rapport à 22°C. Par ailleurs, quelles que soient les conditions thermiques (22°C ou 25°C) on ne constate aucun changement significatif de la fertilité des oeufs pondus par les femelles survivantes. Les auteurs concluent que les effets secondaires de B. bassiana sur la fécondité du doryphore dépendent des conditions thermiques et qu'ils pourraient devenir négligeables à des températures élevées en plein champ.

     

A Sex Chromosome piRNA Promotes Robust Dosage Compensation and Sex Determination in C. elegans

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Behavioral sensitization and tolerance to cocaine and the occupation of dopamine receptors by dopamine

Data from the authors’ laboratory on the neural substrates of Pavlovian conditioning and behavioral sensitization to psychomotor stimulants are reviewed. The findings of a recent experiment on the role of occupation of dopamine receptors by dopamine and its association to behavioral sensitization are reported. Daily intermittent injections of cocaine produced behavioral sensitization to the locomotor response in rats, whereas continuous cocaine infusions produced behavioral tolerance. Behavioral sensitization to cocaine was blocked by coadministration of nimodipine, anL-type calcium channel blocker. The increases in locomotion produced by cocaine was associated with an increase in the occupation of striatal dopamine D₁ and D₂ receptors, measured as the density of receptors protected from denaturation byN-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ). This association was not observed when rats were given a challenge injection of cocaine 10 d after withdrawal from similar treatment regimens. Rats given a cocaine challenge after withdrawal from either intermittent or continuous cocaine treatment regimens exhibited increased occupations of striatal D₁ and D₂ receptors. This increase was similar in magnitude to that observed in rats without a history of cocaine treatments after a challenge injection of cocaine. This suggests tnat the differences in occupancy of striatal dopamine receptors by dopamine observed in the prewithdrawal condition are likely the result of differences in brain levels of cocaine achieved by the two treatment regimens. Occupancy of striatals dopamine D₁ and D₂ receptors does not appear to be related to the development of sensitization to the motor-stimulating effects of cocaine.     

Expression of recombinant human granulocyte colony-stimulating factor in CHO dhfr cells: new insights into the in vitro amplification expression system

The in vitro amplification method for heterologous gene expression in mammalian cells is based on the stable transfection of cells with long, linear DNA molecules having several copies of complete expression units, coding for the gene of interest, linked to one terminal unit, coding for the selectable marker. DNA concatenamers containing additional expression units can also be prepared: we exploited this feature by co-polymerizing expression units coding for granulocyte colony-stimulating factor (G-CSF) with cassettes for dihydrofolate reductase (DHFR) and for neomycin (Nm) resistance, as selectable markers. We were thus able to obtain high level production of G-CSF in chinese hamster ovary (CHO) dhfr^- cells by combining in vitro amplification to just one step of in vivo amplification. This approach required a considerably shorter time than the classical, stepwise amplification by methotrexate.     

酶的基因剂量效应及其对鱼类远缘杂交的影响

亲缘关系较远的杂种二倍体,酶的表达出现父本基因受抑制,双亲基因表达迟缓和母本基因提前表达三种类型的紊乱。由于双亲等位基因不亲和,各等位基因只调控产生1／2的基因产物,从而致使胚胎出现“双单倍体综合症”而在孵化期全部死亡。远缘杂交三倍体由于母本基因剂量加倍,一方面抑制了父本基因的表达,消除了不协调基因活动的干扰,另一方面母本基因产物趋于正常,代谢得以正常进行,杂种后代能成活并正常生长。     

GDP与CHOP方案治疗非特异性外周T 细胞淋巴瘤的临床对比研究

目的:探讨GDP与CHOP方案治疗非特异性外周T细胞淋巴瘤的临床疗效,为临床治疗提供参考。方法:选择2013年1月到2016年1月我院收治的非特异性外周T细胞淋巴瘤患者80例,随机分为GDP组(n=40)和CHOP组(n=40)。GDP组患者给予GDP治疗方案(顺铂+吉西他滨+强的松),CHOP组患者给予CHOP治疗方案(多柔比星+环磷酰胺+长春新碱+强的松),两组患者均治疗6个疗程。比较两组患者临床疗效和不良反应发生情况。结果:GDP组患者近期疗效总有效率为75.00%,明显高于CHOP组的45.00%,差异具有统计学意义(P0.05)。GDP组患者的无进展生存期(PFS)、总生存期(OS)分别为(9.69±1.50)月和(16.72±3.06)月,明显大于CHOP组的(5.16±1.38)月和(10.98±3.37)月,差异具有统计学意义(P0.05)。GDP组患者恶心呕吐的发生率为72.50%,明显低于CHOP组的97.50%,差异具有统计学意义(P0.05)。结论:GDP方案治疗非特异性外周T细胞淋巴瘤的临床疗效明显优于CHOP方案,且不良反应发生率低,值得在临床上推广应用。     

Des dipeptides soufres differencient nettement Vigna radiata de Vigna mungo

The presence of γ-glutamyl-S-methylcysteine and its sulphoxide characterizes the seeds of Vigna radiata; on the other hand, those of Vigna mungo do not contain these compounds but are particularly rich in γ-glutamylmethionine and its sulphoxide. These two peptides, absent in V. radiata, represent an important fraction of the methionine existing in V. mungo and bring the total amount of this amino acid up to a relatively high level. Thereby, they are liable to play an important part in the improvement of the nutritive value of certain legume crops.     

Pathogenicity of entomopathogenic hyphomycetes, Paecilomyces fumoso-roseus and Nomuraea rileyi, to eggs of noctuids, Mamestra brassicae and Spodoptera littoralis

Bioassays were conducted to determine the susceptibility of egg masses of Mamestra brassicae and Spodoptera littoralis to different spore doses of Paecilomyces fumoso-roseus and Nomuraea rileyi at 20° and 25°C. P. fumoso-roseus was highly virulent against eggs, whereas N. rileyi provoked only a deferred mortality of larvae hatched from treated eggs. Nevertheless, larval mortality of S. littoralis caused by N. rileyi at 25°C was more effective after first-instar larval contamination than after egg mass treatment. The duration of the egg stage could explain differences of susceptibility between the two noctuids at 25°C. Scanning electron microscopical observations suggested two ways of contamination of newly hatched larvae. First, fungal germinations on the chorion surface suggested that newly hatched larvae might be infected by penetration of the egg integument before hatching. Second, conidia on the egg cuticle could be an entomopathogenic inoculum for newly emerging larvae which fed upon chorions. Results showed that pathogenicity of Hyphomycetes to noctuid eggs might be a promising area of investigation for biological control.     

EFFECTS OF RAPE GROWN IN SO2-ENRICHED ATMOSPHERES ON PERFORMANCE OF THE APHID,MYZUS PERSICAE (SULZER) *

Abstract Performances of the aphid, Myzus persicae, were monitored during feeding on leaves of the rape, Brassica chinensis, previously fumigated with SO₂ in a range of concentrations between 20 and 160 nl/1 at 25°C. Response of the aphids depended on dose of the air pollutant. Moderate dose of SO₂ treatment on the rape greatly enhanced the aphids' performances while high level of the pollutant heavily suppressed their growth and reproduction. They performed best at about 80 nl/l SO₂, where nymphal mean relative growth rate (MRGR), adult fecundity and intrinsic increase rate(r_m) of the population increased by 31.5%, 89.8% and 42. 7% over those of control aphids, respectively. Determination of the plant chemistry suggests that the altered relative content of methionine induced by So, exposure might be responsible for this response of dosage dependence.