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  • 1
    ISSN: 1432-2145
    Keywords: Key words NTP303 glycoprotein ; Pollen-specific Ntp303 gene ; Pollen-tube specific protein ; Plant reproduction ; Biochemical analysis ; Pollen development
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The pollen-specific gene Ntp303 belongs to the class of late pollen specific genes. It is first transcribed directly after pollen mitosis. Biochemical properties, appearance and precise location of the NTP303 protein during pollen development and pollen tube growth were studied by amino-acid micro-sequencing, protein gel blotting and immuno-localization. Antisera were raised against recombinant proteins, encoded by sequences of the pollen-specific Ntp303 gene. The antibodies specifically recognized a 69-kDa glycoprotein. Electron-microscopic immuno-localization of the protein revealed the presence of high concentrations of the NTP303 protein at the vegetative plasma membranes that surround the vegetative cell, the generative cell and the sperm cells of pollen and pollen tubes. The generative plasma membranes of the generative cell and the sperm cells were negative. NTP303 protein was also present in the cell walls and in callose plugs. With this method it was shown that the NTP303 protein was already present in mid-bicellular pollen, after the first, asymmetrical pollen mitosis. Possible functions for the NTP303 protein are discussed in relation to its properties and its association with the vegetative plasma membranes.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-2145
    Keywords: Key words LIM cDNAs ; Pollen-specific ; Zinc finger ; Nicotiana tabacum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  A PCR-generated DNA probe covering the region corresponding to the two contiguous LIM domain sequences from the sunflower PLIM-1 cDNA was used to screen a cDNA library prepared from poly-A+ RNA isolated from mature Nicotiana tabacum pollen. Two clones were isolated and found to have unique but closely related nucleotide sequences NTPLIM1a and NTPLIM1b. Homology searches revealed the presence of two LIM domain repeats in each clone. Detailed spatial and temporal northern blot analysis showed that the NTPLIM transcripts were present specifically in mature and germinating pollen and were detectable immediately before the asymmetrical division pollen mitosis I, reaching a maximal level in mature pollen. Southern blot analysis revealed the NTPLIM1a and NTPLIM1b genes to be single-copy and members of a small gene family in the N. tabacum genome. Analysis of a genomic clone encoding NTPLIM1a revealed that the gene is split by four introns ranging from 79 to 922 bp in length. The high levels of developmentally regulated expression specifically in pollen suggests an important role for these proteins in pollen maturation and/or function.
    Type of Medium: Electronic Resource
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