Kumar V, Trick M
Cambridge Laboratory, AFRC Institute of Plant Science Research, John Innes Centre, Norwich, UK.
Mol Gen Genet. 1993 Nov;241(3-4):440-6. doi: 10.1007/BF00284698.
A genomic library from an S29/S29 self-incompatible genotype of Brassica oleracea was screened with a probe carrying part of the catalytic domain of a Brassica S-receptor kinase (SRK)-like gene. Six positive phage clones with varying hybridisation intensities (K1 to K6) were purified and characterised. A 650-700 bp region corresponding to the probe was excised from each clone and sequenced. DNA and predicted protein sequence comparisons based on a multiple alignment identified K5 as a pseudogene, whereas the others could encode functional proteins. K3 was found to have lost an intron from its genomic sequence. The six genes display different degrees of sequence similarity and form two distinct clusters in a dendrogram. The 98% similarity between K4 and K6, which extends across intron sequences, suggests that these might be very recently diverged alleles or daughters of a duplication. In addition, K2 showed a comparably high similarity to the probe. Clones K1, K3 and K5 cross-hybridised with an SLG29 cDNA probe, indicating the presence of upstream receptor domains homologous to the Brassica SLG gene. This suggests that the previously reported S sequence complexity may be ascribed to a large receptor kinase gene family.
用携带甘蓝型油菜S受体激酶(SRK)样基因部分催化结构域的探针筛选甘蓝型油菜S29/S29自交不亲和基因型的基因组文库。纯化并鉴定了六个杂交强度不同的阳性噬菌体克隆(K1至K6)。从每个克隆中切下对应于探针的650 - 700 bp区域并进行测序。基于多重比对的DNA和预测蛋白质序列比较确定K5为假基因,而其他克隆可编码功能蛋白。发现K3的基因组序列缺失了一个内含子。这六个基因显示出不同程度的序列相似性,并在系统发育树中形成两个不同的簇。K4和K6之间98%的相似性延伸到内含子序列,表明它们可能是最近分化的等位基因或重复产生的子代。此外,K2与探针显示出相当高的相似性。克隆K1、K3和K5与SLG29 cDNA探针交叉杂交,表明存在与甘蓝型油菜SLG基因同源的上游受体结构域。这表明先前报道的S序列复杂性可能归因于一个大型受体激酶基因家族。