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发育调控的植物基因:一个小麦醇溶蛋白基因组克隆的核苷酸序列

Developmentally regulated plant genes: the nucleotide sequence of a wheat gliadin genomic clone.

作者信息

Rafalski J A, Scheets K, Metzler M, Peterson D M, Hedgcoth C, Söll D G

出版信息

EMBO J. 1984 Jun;3(6):1409-15. doi: 10.1002/j.1460-2075.1984.tb01985.x.

Abstract

Gliadins, the major wheat seed storage proteins, are encoded by a multigene family. Northern blot analysis shows that gliadin genes are transcribed in endosperm tissue into two classes of poly(A)+ mRNA, 1400 bases (class I) and 1600 bases (class II) in length. Using poly(A)+ RNA from developing wheat endosperm we constructed a cDNA library from which a number of clones coding for alpha/beta and gamma gliadins were identified by hybrid-selected mRNA translation and DNA sequencing. These cDNA clones were used as probes for the isolation of genomic gliadin clones from a wheat genomic library. One such genomic clone was characterized in detail and its DNA sequence determined. It contains a gene for a 33-kd alpha/beta gliadin protein (a 20 amino acid signal peptide and a 266 amino acid mature protein) which is very rich in glutamine (33.8%) and proline (15.4%). The gene sequence does not contain introns. A typical eukaryotic promoter sequence is present at -104 (relative to the translation initiation codon) and there are two normal polyadenylation signals 77 and 134 bases downstream from the translation termination codon. The coding sequence contains some internal sequence repetition, and is highly homologous to several alpha/beta gliadin cDNA clones. Homology to a gamma-gliadin cDNA clone is low, and there is no homology with known glutenin or zein cDNA sequences.

摘要

醇溶蛋白是小麦种子的主要贮藏蛋白,由一个多基因家族编码。Northern印迹分析表明,醇溶蛋白基因在胚乳组织中转录成两类多聚腺苷酸化的mRNA,长度分别为1400个碱基(I类)和1600个碱基(II类)。利用发育中小麦胚乳的多聚腺苷酸化RNA,我们构建了一个cDNA文库,通过杂交选择的mRNA翻译和DNA测序,从该文库中鉴定出了一些编码α/β和γ醇溶蛋白的克隆。这些cDNA克隆被用作探针,从一个小麦基因组文库中分离基因组醇溶蛋白克隆。对其中一个这样的基因组克隆进行了详细表征,并测定了其DNA序列。它包含一个编码33-kdα/β醇溶蛋白的基因(一个20个氨基酸的信号肽和一个266个氨基酸的成熟蛋白),该蛋白富含谷氨酰胺(33.8%)和脯氨酸(15.4%)。该基因序列不含内含子。在-104(相对于翻译起始密码子)处存在一个典型的真核启动子序列,在翻译终止密码子下游77和134个碱基处有两个正常的多聚腺苷酸化信号。编码序列包含一些内部序列重复,并且与几个α/β醇溶蛋白cDNA克隆高度同源。与γ醇溶蛋白cDNA克隆的同源性较低,与已知的麦谷蛋白或玉米醇溶蛋白cDNA序列没有同源性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fd2b/557531/4582629d362e/emboj00310-0190-a.jpg

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