Sun C, Sathish P, Ahlandsberg S, Jansson C
Department of Biochemistry, The Arrhenius Laboratories, Stockholm University, S-106 91 Stockholm, Sweden.
Plant Physiol. 1998 Sep;118(1):37-49. doi: 10.1104/pp.118.1.37.
The sbeIIa and sbeIIb genes, encoding starch-branching enzyme (SBE) IIa and SBEIIb in barley (Hordeum vulgare L.), have been isolated. The 5' portions of the two genes are strongly divergent, primarily due to the 2064-nucleotide-long intron 2 in sbeIIb. The sequence of this intron shows that it contains a retro-transposon-like element. Expression of sbeIIb but not sbeIIa was found to be endosperm specific. The temporal expression patterns for sbeIIa and sbeIIb were similar and peaked around 12 d after pollination. DNA gel-blot analysis demonstrated that sbeIIa and sbeIIb are both single-copy genes in the barley genome. By fluorescence in situ hybridization, the sbeIIa and sbeIIb genes were mapped to chromosomes 2 and 5, respectively. The cDNA clones for SBEIIa and SBEIIb were isolated and sequenced. The amino acid sequences of SBEIIa and SBEIIb were almost 80% identical. The major structural difference between the two enzymes was the presence of a 94-amino acid N-terminal extension in the SBEIIb precursor. The (beta/alpha)8-barrel topology of the alpha-amylase superfamily and the catalytic residues implicated in branching enzymes are conserved in both barley enzymes.
已分离出编码大麦(Hordeum vulgare L.)中淀粉分支酶(SBE)IIa和SBEIIb的sbeIIa和sbeIIb基因。这两个基因的5'部分差异很大,主要是由于sbeIIb中有2064个核苷酸长的内含子2。该内含子的序列表明它含有一个类似反转录转座子的元件。发现sbeIIb而非sbeIIa的表达具有胚乳特异性。sbeIIa和sbeIIb的时间表达模式相似,在授粉后约12天达到峰值。DNA凝胶印迹分析表明,sbeIIa和sbeIIb在大麦基因组中均为单拷贝基因。通过荧光原位杂交,sbeIIa和sbeIIb基因分别定位于第2和第5号染色体上。分离并测序了SBEIIa和SBEIIb的cDNA克隆。SBEIIa和SBEIIb的氨基酸序列几乎80%相同。这两种酶之间的主要结构差异是SBEIIb前体中存在一个94个氨基酸的N端延伸。α-淀粉酶超家族的(β/α)8桶拓扑结构以及与分支酶相关的催化残基在两种大麦酶中均保守。