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构建了日本梨树(Pyruspyrifolia)花粉cDNA文库, 发现了一个编码花粉管分泌的类β-1,3-葡聚糖酶蛋白(BGN-1)的cDNA(bgn-1 )并测定了其序列。bgn-1由1408 bp组成, 包括47 bp的5’-非翻译区, 由1194 bp组成并编码了397个氨基酸的ORF和167 bp的3’-非翻译区。3’-非翻译区含有1个NUE(AATTAA)和3个似FUE的基序, 分别位于1369~1374(AAT-TAA)、1320~1325、1327~1332(TTTTTA)和1363~1368(TTTGGA)。 bgn-1编码的蛋白(BGN-1)与DDBJ中的植物β-1,3-葡聚糖酶的序列相似性范围为37%~59%, 与3D结构已知的大麦β-1, 3-葡聚糖酶(GⅡ)的序列相似性为39.7%。对GⅡ与BGN-1进行的疏水簇分析(HCA同源分为87.1%, >75%的一般标准)和使用3D-PSSM软件进行的分析暗示, BGN-1的3D结构与大麦GⅡ同源性最大(≧95%的肯定度)。BGN-1的信号肽长度为22 aa, 成熟的BGN-1(375 aa)的理论分子量为4 0723 D, 理论pI值为9.59, 诊断氨基酸残基模式(D, L, S, L)与禾谷类的与生长相关的亚家族D的诊断氨基酸残基模式(D, L, S, Q)极相似。在BGN-1的C-端延伸区的位点352~367之间发现了1个功能不清, 重复出现的ProXXPro结构。
A cDNA library of Japanese Pyrus pyrifolia pollen was constructed and a cDNA (bgn-1) encoding β1,3-glucanase secreted by pollen tubes (BGN-1) was found and sequenced. The bgn-1 consists of 1408 bp, including a 47 bp 5’-untranslated region consisting of 1194 bp and encoding a 397 amino acid ORF and a 167 bp 3’-untranslated region. The 3’-untranslated region contains one NUE (AATTAA) and three FUE-like motifs located at 1369-1374 (AAT-TAA), 1320-1325,1327-1332 (TTTTTA) and 1363-1368 (TTTGGA) . The sequence similarity between bgn-1 protein (BGN-1) and plant β-1,3-glucanase in DDBJ ranged from 37% to 59%, which was in agreement with that of barley β-1, 3 - glucanase (G Ⅱ) sequence similarity of 39.7%. Hydrophobic cluster analysis of GII with BGN-1 (HCA homology divided into 87.1%,> 75% of the general standard) and analysis using 3D-PSSM software suggested that the 3D structure of BGN-1 is homologous to the GII of barley Maximum (≧ 95% affirmative). The signal peptide length of BGN-1 was 22 aa. The theoretical molecular weight of mature BGN-1 (375 aa) was 40 0723 D and the theoretical pI value was 9.59. The amino acid residue pattern (D, L, S, L) The diagnostic amino acid residue patterns (D, L, S, Q) of the growth-related subfamily D in cereals are very similar. One of the unclear, recurring ProXXPro constructs was found between positions 352 and 367 of the C-terminal extension of BGN-1.