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本实验以12个3’锚锭引物和10个5’随机引物构成引物组合,运用mRNA差异显示方法比较玉米品种“农大3138”萌发48h胚芽和胚根的基因表达差异。结果表明,在120个引物组合中有40个引物组合可以扩增出差异条带,其中以AG、GA、GG、AT结尾的锚锭引物扩增效果较好。对经反向Northern验证的6个片段克隆测序,并在Genebank中进行同源性比较,发现2个在胚芽和胚报中表达量都很高的cDNA片段是编码玉米β-D-葡糖苷酶的基因。一个在胚芽中表达而在胚根中不表达的片段是编码线粒体分子伴侣的基因。另外3个在胚芽中表达量高于在胚根中表达的片段未查到同源序列,有可能是新基因。
In this experiment, 12 3 ’anchor primers and 10 5’ random primers were used to construct the primer combinations. MRNA differential display was used to compare the gene expression differences of germ and radicle at 48 h germination of maize variety “Nongda 3138”. The results showed that 40 primer combinations could amplify the differential bands. Among them, the anchor primers ending with AG, GA, GG and AT had a good amplification effect. The 6 fragments verified by reverse Northern blotting were cloned and compared in Genebank for homology. The two cDNA fragments which were highly expressed in embryo and embryo were found to encode maize β-D-glucosidase Of the gene. A fragment that is expressed in the germ but not expressed in the radicle is the gene that encodes the chaperone. The other three genes whose expression level was higher in germ than those expressed in radicle did not find homologous sequences, which may be new genes.