Anderson M A, McFadden G I, Bernatzky R, Atkinson A, Orpin T, Dedman H, Tregear G, Fernley R, Clarke A E
Plant Cell Biology Research Centre, School of Botany, University of Melbourne, Parkville, Victoria, Australia.
Plant Cell. 1989 May;1(5):483-91. doi: 10.1105/tpc.1.5.483.
Three alleles of the self-incompatibility gene of Nicotiana alata have been cloned and sequenced. A comparison of the sequences shows a surprisingly low level of homology (56%) and the presence of defined regions of homology and variability. The homologous regions include the N-terminal sequence, most of the cysteine residues and glycosylation sites, as well as other blocks throughout the sequence. We interpret these conserved regions as "framework" and nonconserved regions as "hypervariable," following the terminology used to describe analogous regions in the IgG supergene family. The low level of overall homology forms the basis of a general method for isolating S-allele-specific cDNAs. Allele-specific antibodies can be generated using synthetic peptides corresponding to one of the variable regions. When applied to sections of the pistil, these antibodies label the intercellular matrix in the stigma and transmitting tissue of the style and the cell walls in the epidermis of the placenta. HindIII digestion of genomic DNA generates a characteristic pattern of S-gene fragments for each genotype. These restriction fragment length polymorphisms can be used to assign S-genotype to progeny arising from breeding experiments.
已克隆并测序了烟草的自交不亲和基因的三个等位基因。序列比较显示同源性水平出奇的低(56%),且存在特定的同源区域和可变区域。同源区域包括N端序列、大部分半胱氨酸残基和糖基化位点,以及整个序列中的其他片段。按照用于描述IgG超基因家族中类似区域的术语,我们将这些保守区域解释为“框架”,非保守区域解释为“高变区”。总体同源性水平较低构成了分离S等位基因特异性cDNA的通用方法的基础。可以使用对应于可变区之一的合成肽产生等位基因特异性抗体。将这些抗体应用于雌蕊切片时,它们会标记柱头中的细胞间基质、花柱的传递组织以及胎座表皮中的细胞壁。基因组DNA经HindIII消化后,每种基因型都会产生S基因片段的特征性图谱。这些限制性片段长度多态性可用于确定育种实验产生的后代的S基因型。