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  • 1995-1999
  • 1990-1994  (2)
  • 1975-1979
  • 1991  (2)
  • Brassica napus  (1)
  • Zona pellucida  (1)
  • 1
    ISSN: 1573-5028
    Keywords: acyl carrier protein ; Brassica napus ; enoyl-ACP reductase ; fatty acid synthesis ; seed development ; nuclear-encoded chloroplast proteins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The onset of storage lipid biosynthesis during seed development in the oilseed crop Brassica napus (rape seed) coincides with a drastic qualitative and quantitative change in fatty acid composition. During this phase of storage lipid biosynthesis, the enzyme activities of the individual components of the fatty acid synthase system increase rapidly. We describe a rapid and simple purification procedure for the plastidlocalized NADH-dependent enoyl-acyl carrier protein reductase from developing B. napus seed, based on its affinity towards the acyl carrier protein (ACP). The purified protein was N-terminally sequenced and used to raise a potent antibody preparation. Immuno-screening of a seed-specific λgt11 cDNA expression library resulted in the isolation of enoyl-ACP reductase cDNA clones. DNA sequence analysis of an apparently full-length cDNA clone revealed that the enoyl-ACP reductase mRNA is translated into a precursor protein with a putative 73 amino acid leader sequence which is removed during the translocation of the protein through the plastid membrane. Expression studies in Escherichia coli demonstrated that the full-length cDNA clone encodes the authentic B. napus NADH-dependent enoyl-ACP reductase. Characterization of the enoyl-ACP reductase genes by Southern blotting shows that the allo-tetraploid B. napus contains two pairs of related enoyl-ACP reductase genes derived from the two distinct genes found in both its ancestors, Brassica oleracea and B. campestris. Northern blot analysis of enoyl-ACP reductase mRNA steady-state levels during seed development suggests that the increase in enzyme activity during the phase of storage lipid accumulation is regulated at the level of gene expression.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 30 (1991), S. 100-104 
    ISSN: 1040-452X
    Keywords: Embryo ; Incision ; Zona pellucida ; Trophoblast ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Experiment I was designed to determine if cell-free synchronous uterine flushings contain an embryotoxic substance that is normally screened by the intact zona pellucida. Sixty 4-cell embryos were allocated to three treatment groups: (1) control embryos (n = 20) were cultured in Modified Kreb's Ringer Bicarbonate medium + 10% bovine calf serum (mKRB-BCS), (2) UF embryos (n = 20) were cultured in 80% mKRB-BCS + 20% sterile dialyzed uterine flushings (UF), (3) MicroUF embryos (n = 20) received a microsurgical incision in the zona pellucida and were cultured in 80% mKRB-BCS + 20% UF. Following 72 h in culture at 37°C under a 90% N2, 5% CO2, and 5% O2 atmosphere, the number of nuclei/embryo and the incidence of protrusion of the trophoblast through the zona pellucida (PTZ) were recorded. Addition of UF had no effect on embryo development. A greater (P 〉 .005) proportion of MicroUF embryos exhibited PTZ as compared to UF and control embryos. Experiment II was devised to further characterize the occurrence of PTZ in Micro porcine embryos. Thirty-three 4- to 10-cell embryos and 14 morulae were distributed across two treatments: (1) control embryos (n = 16 and 6, respectively) were cultured as described in Experiment I; and (2) micro embryos were treated similarly to MicroUF embryos in Experiment I but were cultured in mKRB-BCS only. At the onset of PTZ, embryos were immediately fixed and examined. The proportion of embryos exhibiting PTZ was greater (P 〈 .007) for Micro versus control embryos. Both the mean time to PTZ and the number of nuclei present were greater for blastocysts cultured from control as opposed to Micro morulae (P 〈 .0001 and P 〈 .0002, respectively). These results indicate that (a) cell-free synchronous uterine flushings are not detrimental to embryos with incised zonae and (b) microsurgery increases the incidence of and reduces the time to protrusion of the embryo through the zona pellucida in cultured embryos.
    Additional Material: 5 Tab.
    Type of Medium: Electronic Resource
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