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为了挖掘和有效利用陕253中的优良基因,根据LMW-GS编码基因5′和3′端的保守序列设计引物,采用PCR方法以陕253的基因组DNA为模板进行扩增,分别回收并克隆到载体上,测序后得到11个新LMW-GS基因。GenBank登录号分别为GQ892576~GQ892580和GQ892583~GQ892588,其长度为903~1 081 bp。其中,GQ892576、GQ892585、GQ892586、GQ892587和GQ892588具有完整编码区,可分别编码305、351、298、351和307个氨基酸残基的成熟蛋白。而GQ892577、GQ892578、GQ892579、GQ892580、GQ892583和GQ892584由于在编码区内出现提前终止密码子,推测为假基因。推导的氨基酸序列结果显示:它们都具有LMW-m型低分子量谷蛋白亚基的典型结构特征。
In order to excavate and effectively use the good genes in Shaan 253, primers were designed according to the conserved sequences at the 5 ’and 3’ ends of the LMW-GS encoding gene, and PCR was performed using genomic DNA of Shaan 253 as a template for amplification and recovery respectively After sequencing, 11 new LMW-GS genes were obtained. The GenBank accession numbers were GQ892576 ~ GQ892580 and GQ892583 ~ GQ892588 respectively, and their lengths ranged from 903 to 1 081 bp. Among them, GQ892576, GQ892585, GQ892586, GQ892587 and GQ892588 have complete coding regions and can encode mature proteins of 305, 351, 298, 351 and 307 amino acid residues respectively. However, GQ892577, GQ892578, GQ892579, GQ892580, GQ892583 and GQ892584 were presumed to be pseudogenes due to the presence of early stop codons in the coding region. The deduced amino acid sequence results showed that they all have the typical structural features of LMW-m low molecular weight glutenin subunits.