Nucleotide Sequence of the [alpha]-Amylase Gene from Vigna mungo
Nucleotide Sequence of the [alpha]-Amylase Gene from Vigna mungo
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来自绿豆的α-淀粉酶基因的核苷酸序列
DOI：
10.1104/pp.103.4.1459
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发表时间：
1993
期刊：
影响因子：
--
通讯作者：
T. Minamikawa
中科院分区：
文献类型：
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作者：
H. Takeuchi;D. Yamauchi;S. Wada;T. Minamikawa
绿豆发芽种子的子叶中α-淀粉酶的量增加（Minamikawa 等，1992）。为了检查该基因的基因组组织，通过 Southem 杂交与 32P 标记的 α-淀粉酶 cDNA 分析了 V. mungo 的 DNA（Yamauchi 和 Minamikawa，1990）。以 kb 为单位的片段大小的近似估计为：EcoRI，8 和 2.2；印度三世，15；和XbnI，4。由于HindIII和XbaI不切割cDNA，结果表明存在单拷贝的α-淀粉酶基因。在通过1％琼脂糖凝胶电泳分离经EcoRI消化的V.mungo的DNA后，将从凝胶切下的约2.2-kb片段连接至Xgt10。我们从该文库中分离出一个基因组克隆 (A VMAMYIA)，其中包含 1499 bp 转录区域和 639 bp 5’侧翼区域。通过聚合酶链式反应分离转录区域的其余部分。扩增并亚克隆 1.4 kb 片段。该克隆被命名为pVMAMY1B，包括第三外显子、第三内含子和第四外显子的一部分，表明来自V.mungo的淀粉酶基因具有235、421和637bp的三个内含子，分隔四个113、130、806和492bp的外显子。内含子的位点与单子叶植物α-淀粉酶基因亚家族2的位点相同(Huang等，1992)。
The amount of a-amylase increased in cotyledons of germinating seeds of Vigna mungo (Minamikawa et al., 1992). To examine the genomic organization of the gene, DNA from V. mungo was analyzed by Southem hybridization with 32Plabeled a-amylase cDNA (Yamauchi and Minamikawa, 1990). Approximate estimates of the fragment sizes in kb were: EcoRI, 8 and 2.2; HindIII, 15; and XbnI, 4. Since HindIII and XbaI do not cut the cDNA, the results indicate that there is a single copy of the a-amylase gene. After the DNA from V. mungo digested by EcoRI was separated by 1% agarose gel electrophoresis, about 2.2-kb fragments cut out from the gel were ligated to XgtlO. From this library we isolated a genomic clone (A VMAMYIA) containing a 1499-bp transcribed region and a 639-bp 5’-flanking region. The rest of the transcribed region was isolated by polymerase chain reaction. A 1.4-kb fragment was amplified and subcloned. This clone, designated pVMAMYlB, included a part of the third exon, the third intron, and the fourth exon, indicating that the aamylase gene from V. mungo has three introns of 235, 421, and 637 bp separating four exons of 113, 130, 806, and 492 bp. The sites of the introns are the same as those of the monocot a-amylase gene subfamily 2 (Huang et al., 1992).