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  • 1
    ISSN: 1432-2145
    Keywords: Confocal laser scanning microscopy ; In situ hybridization ; Nicotiana tabacum ; Pollen-specific gene ; Promoter analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The localization of transcripts of the pollenspecific gene NTP303 has been determined by means of in situ hybridization in combination with confocal laser scanning microscopy. NTP303 transcripts were first detectable at the mid-bi-nucleate stage of pollen development and persisted even after the pollen tube had reached the ovary. The majority of the NTP303 transcripts were localized in the vegetative cell, and were predominantly present in the apex of the pollen tube. To study the mechanism of regulation, the genomic clone NTPg303 was isolated and characterized. NTPg303 belongs to a gene family of at least five members. No introns are present in its coding region. Regions matching the pollen-specific PB-element TGTGGTT (Twell et al. 1991) have been found.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1573-5028
    Keywords: anther ; in situ hybridization ; microspore development ; microspore-specific gene ; Nicotiana tabacum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The characterization of a gene with a unique microspore-specific expression pattern is reported. Isolated microspores from tobacco were used to synthesize a cDNA library. Clones that did not hybridize to leaf cDNA were further characterized by northern analysis. One clone proved to be a microspore-specific cDNA, representing a transcript of 650 nt. The corresponding gene, NTM19 (Nicotiana tabacum microspore-specific), was isolated and its sequence analysed. The gene encodes a protein of 10.8 kDa with a pI of 6.92 and a putative signal sequence at the N-terminus. A localization study revealed a unique spatial and temporal distribution. The transcript was only detected in the unicellular microspore. No hybridization signals were observed in other pollen developmental stages, nor in the surrounding anther tissues or other vegetative tissues of the plant. Therefore it can be concluded that NTM19 is a gene with a highly microspore-specific character according to both localization and stage of expression. Southern blot analysis demonstrated the presence of a small gene family. The occurrence of TNM19 was investigated in a range of closely and distantly related species and was found to be present in other solanaceous species, including the ancestors of tobacco and in a monocot species.
    Type of Medium: Electronic Resource
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