Plant Breeding Institute, CB2 2LQ, Cambridge, UK.
Plant Mol Biol. 1985 Jan;5(1):13-24. doi: 10.1007/BF00017869.
The α-Amy1 and α-Amy2 genes of wheat produce distinct subsets of α-amylase isozymes which show different patterns of expression in wheat aleurone cells and in developing grain. In order to characterise the organisation and expression of these genes, clones of α-Amy1 and α-Amy2 cDNA have been isolated. The two types of cDNA clone were distinguished within a small library of α-amylase cDNA clones (Baulcombe and Buffard, Planta 157 493-501 [1983]) by restriction endonuclease mapping and by cross hybridisation. The identity of α-Amy1 or α-Amy2 type was assigned from the results of hybrid selected translation analysis in which small subfragments of the cDNA clones were used. These subfragments were derived from the 3' ends of the cDNA and did not cross hybridise between the different types of cDNA. Hybridisation of α-Amy1 and α-Amy2 cDNA probes to restriction enzyme digests of wheat nuclear DNA revealed that these are multigene families located on the group 6 (α-Amy1) and group 7 (α-Amy2) chromosomes. Studies on the levels of α-Amy1 and α-Amy2 mRNA in developing grain and in aleurone tissue indicated that the differences in isozyme expression are due to the patterns of mRNA accumulation. In aleurone tissue the α-Amy1 transcripts accumulate in parallel with other genes which are regulated by gibberellic acid, while the accumulation of α-Amy2 genes is sustained for 36 h longer.
小麦的α-Amy1 和 α-Amy2 基因产生不同的α-淀粉酶同工酶亚基,这些同工酶在小麦糊粉层细胞和发育中的谷物中表现出不同的表达模式。为了描述这些基因的组织和表达,我们分离了α-Amy1 和 α-Amy2 cDNA 的克隆。通过限制内切酶图谱分析和交叉杂交,在一个小的α-淀粉酶 cDNA 克隆文库(Baulcombe 和 Buffard,Planta 157 493-501 [1983])中区分了这两种类型的 cDNA 克隆。通过杂交选择翻译分析,从小 cDNA 克隆的 3' 末端获得的亚片段,确定了 α-Amy1 或 α-Amy2 型的身份。这些亚片段来源于 cDNA 的 3' 末端,并且在不同类型的 cDNA 之间不发生交叉杂交。α-Amy1 和 α-Amy2 cDNA 探针与小麦核 DNA 的限制酶消化产物的杂交表明,这些基因是位于第 6 组(α-Amy1)和第 7 组(α-Amy2)染色体上的多基因家族。对发育中的谷物和糊粉层组织中 α-Amy1 和 α-Amy2 mRNA 水平的研究表明,同工酶表达的差异是由于 mRNA 积累模式的不同。在糊粉层组织中,α-Amy1 转录物与其他受赤霉素调节的基因平行积累,而α-Amy2 基因的积累则持续 36 小时。