摘要
采用PCR技术,分别从马铃薯、金花菊基因组DNA中扩增到了马铃薯损伤诱导启动子Wun 1,和对启动子活性有显著增强作用的Me1 3 非转录区。经序列同源性分析表明,马铃薯损伤诱导启动子Wun 1与已知序列的同源性为96.86 %,Me1 3 非转录区的同源性达99 %。得到的Wun 1克隆与原序列有一定的差异,尤其在735～768处差异较大,为一新颖的启动子,现已注册到分子生物学GenBank数据库(GenBank, gi: AY485646)。利用来自欧文氏梨火疫病菌的Hrp基因与克隆的Wun 1启动子和Me1 3 非转录区构建了Wun 1-Hrp-Me1 3 植物表达载体pBI WHM,同时构建了CaMV 35S-Hrp-Me1 3 植物表达载体pBI HM,为下一步鉴定Wun 1启动子和Me1 3 增强子对Hrp基因表达活性的影响及通过遗传转化培育植物抗病品种奠定了基础。
The potato wound-inducible promoter Wun 1 gene, Me1 3 untranslated region which has notable enhance function to promoter action of were amplified from Solanum tuberosum and Flaveria bidentis respectively,with polymerase chain reaction (PCR). Sequence comparison showed that potato wound-inducible promoter Wun 1 genes were homologious by 96.86 % to the corresponding sequences published on GenBank, Me1 3 untranslated region by 99 %. In view of difference in certain extent between the cloned fragment sequences in the test and the corresponding sequences published on GenBank,significant especially at those loci from 735 to 768, promoter Wun 1 gene sequence have been registered on GenBank (gi:AY485646). Two of plant expression, pBIWHM (Wun 1-Hrp-Me1 3) and pBIHM (35S-Hrp-Me1 3) , have been constructed,thus, laying a foundation for identifying the effects of promoter Wun1 and Me1 3 untranslated region to Erwinia amylovora Hrp gene and breeding varieties of potato with better disease resistance by genetic transformation.
出处
《甘肃农业大学学报》
CAS
CSCD
2004年第2期124-130,共7页
Journal of Gansu Agricultural University
基金
国家高技术研究发展计划"863"项目(编号:2001AA241132)
