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首页> 外文期刊>Protoplasma: An International Journal of Cell Biology >Isolation and characterisation of two wheat beta-expansin genes expressed during male gametophyte development
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Isolation and characterisation of two wheat beta-expansin genes expressed during male gametophyte development

机译：男性配子体发育过程中表达的两个小麦β-扩张蛋白基因的分离与鉴定

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摘要

Two novel beta-expansin genes, TaEXPB1 and TaEXPB2, were isolated from wheat microspores by suppression subtractive hybridisation. Northern blot and reverse transcription PCR analyses showed that the expression of both genes was restricted to early stages of male gametophyte development (from microspores to immature pollen). A homology search showed high similarity of the newly discovered genes to generative beta-expansins in grass pollen (group 1 pollen allergens). Southern hybridisation revealed that the isolated genes belong to a distinct group within the subfamily of beta-expansin genes in the wheat genome. A comparison of full-length cDNAs with the corresponding genomic sequences showed that there are two introns in the TaEXPB1 gene, whereas TaEXPB2 has three introns. Both genes were predicted to encode highly similar basic proteins (pI 9.0) with molecular masses of approximately 29 kDa consisting of a signal peptide, catalytic, and polysaccharide binding domains, which include conserved cysteines and tryptophans and motifs characteristic for beta-expansins.
机译：通过抑制性消减杂交，从小麦小孢子中分离出两个新的β-expansin基因TaEXPB1和TaEXPB2。 Northern印迹和逆转录PCR分析表明，这两个基因的表达都限于雄配子体发育的早期阶段（从小孢子到未成熟花粉）。同源性搜索显示，新发现的基因与草花粉（第1组花粉过敏原）中生成的β-扩展酶高度相似。 Southern杂交揭示了分离的基因属于小麦基因组中β-扩张蛋白基因的亚家族中的不同组。全长cDNA与相应基因组序列的比较表明，TaEXPB1基因中有两个内含子，而TaEXPB2基因中有三个内含子。预测这两个基因编码的分子质量约为29 kDa的高度相似的基本蛋白（pI 9.0），由信号肽，催化和多糖结合域组成，其中包括保守的半胱氨酸和色氨酸以及β-扩展蛋白的特征性基序。

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