编码日本梨树(Pyruspyrifolia )花粉类β-1,3-葡聚糖酶蛋白的cDNA克隆与核苷酸序列(英文)  

Cloning and Nucleotide Sequence of the cDNA Encoding a β-1,3-Glucanase-like Protein Secreted from Growing Pollen Tubes of Japanese Pear (Pyrus pyrifolia)

作　　者：周跃钢[1] Naoko Norioka 李绍良[2] Shigemi Norioka 

机构地区：[1]重庆工学院生物工程学院 [2]日本大阪大学蛋白质研究所

出　　处：《农业生物技术学报》2004年第4期374-379,共6页Journal of Agricultural Biotechnology

基　　金：Supported by China Scholarship Council scholarship (No. 98326100) ; a Grant-in-Aid for Special Research on Priority Areas (B) (11238204) from the Ministry of Education, Science, Sports and Culture, Japan.

摘　　要：构建了日本梨树(Pyrus pyrifolia)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(bgm-I)许测定了其序列：bgn-I由1408bp组成，包括47bp的5’-非翻译区，由1194bp组成并编码了397个氨基酸的ORF和167bp的3’-非翻译区。3’-非翻译区含有1个NLTE(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-I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22aa，成熟的BGN—1(375aa)的理论分子量为40723D，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 pollen cDNA library of Japanese pear (Pyrus pyrifolia ) of the family Rosaceae was constructed and a cDNA (bgn-1) of 1 408 bp, which encodes a protein (BGN-1) secreted from growing pollen tubes, has been cloned and sequenced. bgn-1 cDNA is composed of a 5'-untranslated region of 47 bp, an open reading frame of 1 194 bp encoding 397 amino acid residues and a complete 3'-untranslated region of 167 bp. Alignment of the deduced amino acid sequence of bgn-1 with that of the barley (Hordeum vulgare )β-1,3-glucanase (GⅡ) showed 39.7% of amino acid identity. Several residues that were critical for the function of GⅡ were conserved in the deduced BGN-1 polypeptide. Moreover, a hydrophobic cluster analysis (HCA) showed an overall HCA homology score of 87.1% and analysis of BGN-1 with the 3D-PSSM program predicted a 3D structure of BGN-1 highly homologous to that of barley GⅡ with ≥95% certainty. These results suggest that the cloned bgn-1 cDNA encode a β-1,3-glucanase-like protein in Japanese pear. The predicted mature protein (375 aa) has a theoretical molecular mass of 40 723 D, a basic pI of 9.59, a diagnostic amino acid residue mode of D, L, S and L which is very similar to that of growth-related subfamily D (D, L, S and Q) in cereal. A ProXXPro repeat is also found between the position 352 and 367 in the C-extension region.

关 键 词：日本梨树 β-1 CDNA克隆 酶蛋白 核苷酸序列 编码 3-葡聚糖酶 序列相似性 氨基酸残基 粉类 cDNA文库 非翻译区 3D结构 生长相关 花粉管 簇分析 同源性 信号肽 分子量 pI值 禾谷类 组成 大麦 模式 诊断 位点 

分 类 号：S661.2[农业科学—果树学] S641.2[农业科学—园艺学]