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Cloning of cDNA and chromosomal location of genes encoding the three types of subunits of the wheat tetrameric inhibitor of insect a-amylase

机译:小麦α-淀粉酶四聚体抑制剂三种亚基编码基因的cDNA克隆和染色体的染色体定位

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摘要

We have characterized three cDNA clones corresponding to proteins CM1, CM3 and CM16, which represent the three types of subunits of the wheat tetrameric inhibitor of insect -amylases. The deduced amino acid sequences of the mature polypeptides are homologous to those of the dimeric and monomeric -amylase inhibitors and of the trypsin inhibitors. The mature polypeptides are preceded by typical signal peptides. Southern blot analysis of appropriate aneuploids, using the cloned cDNAs as probes, has revealed the location of genes for subunits of the CM3 and of the CM16 type within a few kb of each other in chromosomes 4A, 4B and 4D, and those for the CM1 type of subunit in chromosomes 7A, 7B and 7D. Known subunits of the tetrameric inhibitor corresponding to genes from the B and D genomes have been previously characterized. No proteins of this class have been found to be encoded by the A genome in hexaploid wheat (genomes AA, BB, DD) or in diploid wheats (AA) and no anti -amylase activity has been detected in the latter, so that the A-genome genes must be either silent (pseudogenes) or expressed at a much lower level.
机译:我们已经表征了对应于蛋白质CM1,CM3和CM16的三个cDNA克隆,它们代表了昆虫淀粉酶小麦四聚体抑制剂的三种亚基。推导的成熟多肽的氨基酸序列与二聚和单体淀粉酶抑制剂以及胰蛋白酶抑制剂的氨基酸序列同源。成熟的多肽之前是典型的信号肽。使用克隆的cDNA作为探针,对适当的非整倍体进行Southern印迹分析,揭示了CM3和CM16类型的亚基的基因在4A,4B和4D染色体以及CM1的彼此之间的几个kb内的位置染色体7A,7B和7D中的亚基类型。先前已经表征了对应于来自B和D基因组的基因的四聚体抑制剂的已知亚基。在六倍体小麦(基因组AA,BB,DD)或二倍体小麦(AA)中,没有发现此类蛋白质由A基因组编码,并且在后者中未检测到抗淀粉酶活性,因此A -基因组基因必须是沉默的(假基因)或以低得多的水平表达。

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