Broothaerts W, Janssens G A, Proost P, Broekaert W F
F.A. Janssens Laboratory of Genetics, Katholieke Universiteit Leuven, Belgium.
Plant Mol Biol. 1995 Feb;27(3):499-511. doi: 10.1007/BF00019317.
Complementary DNA clones representing two alleles of the self-incompatibility (S) locus of apple (Malus x domestica Borkh.) have been isolated and characterised. One of the alleles corresponds to a 29 kDa ribonuclease (S-RNase) that was purified from pistil tissue. On northern blots, both cDNAs hybridized to a transcript that was only present in pistils and not in the other plant tissues analysed. Corresponding genomic sequences, amplified by PCR, were found to contain a single intron of 138 bp and 1100 bp respectively. Comparison of both sequences shows that the cDNAs encode mature proteins containing 65% of identical residues. Eight invariable cysteine residues, conserved regions around two histidines thought to play a role in RNA catalysis, and a number of other distinct residues are conserved between the apple S-RNases and similar proteins in the family Solanaceae. As this is the first report of sequences of S-alleles from a species belonging to a family that is not related with the Solanaceae, the structural features of S-RNases deduced from a comparison of their sequences are discussed.
已分离并鉴定了代表苹果(Malus x domestica Borkh.)自交不亲和性(S)位点两个等位基因的互补DNA克隆。其中一个等位基因对应于一种从雌蕊组织中纯化得到的29 kDa核糖核酸酶(S-RNase)。在Northern印迹上,两个cDNA均与仅存在于雌蕊中而不存在于所分析的其他植物组织中的转录本杂交。通过PCR扩增得到的相应基因组序列分别含有138 bp和1100 bp的单个内含子。两种序列的比较表明,cDNA编码的成熟蛋白含有65%的相同残基。苹果S-RNases与茄科中的类似蛋白之间,有八个不变的半胱氨酸残基、两个被认为在RNA催化中起作用的组氨酸周围的保守区域以及许多其他不同的残基是保守的。由于这是来自一个与茄科无关的科的物种的S等位基因序列的首次报道,因此讨论了通过比较其序列推导得出的S-RNases的结构特征。