摘要
应用同源序列克隆策略从铝盐胁迫处理的甜瓜属野生种(Cucumis hystrix Chakr.)叶片中获得了612bp的片段,通过在EST数据库进行同源检索,发现一条甜瓜EST序列(GenBank登录号:AM724963.2)与之高度同源,据此设计引物并经RT-PCR扩增和序列拼接,最后获得了铝诱导基因cDNA序列全长,命名为ChAI(GenBank登录号:FJ968438)。序列分析表明该基因全长967bp,其中开放读码框(ORF)长711bp编码236个氨基酸组成的多肽,5′和3′端非编码区长度各为15bp和241bp。DNAMAN同源性分析表明该基因与葡萄、棉花、羊乳(轮叶党参)、白骨壤(真红树植物)、拟南芥等植物的铝诱导蛋白的氨基酸序列具有78.6%以上的同源性。运用半定量RT-PCR技术对ChAI基因的转录水平进行分析,根据该基因在铝胁迫下大量表达推测ChAI与耐铝盐胁迫响应相关,但具体耐铝胁迫机制还有待深入研究。
Degenerated primers were designed based on conserved amino acids region of aluminum-induced protein from other plants,and a 612bp special fragment was amplified from the wild Cucumis species（Cucumis hystrix Chakr.） leaves under aluminum stress through homologous strategy.Using this fragment sequence data as a querying probe,one highly homologous melon EST sequence（Accession： AM724963.2） was detected.Primers designed based on this EST sequence,the full-length of aluminum-induced gene,designated as ChAI（GenBank Accession no： FJ968438）,was obtained by RT-PCR method and sequence assembling.Sequence analysis showed that ChAI was 967bp in length,which encompassed an 711bp open reading frame（ORF） encoding 236 amino acid residues,and adjacent non-translation 15bp of 5′-terminal region and 241bp of 3′-terminal region,respectively.The results of DNAMAN homologous analysis demonstrated that ChAI had more than 78.6% identity of the amino acids residue with Vitis vinifera,Gossypium hirsutum,Codonopsis lanceolata,Avicennia marina and Arabidopsis thaliana.The transcription analysis of ChAI by semi-quantitative-PCR revealed the high expression under Al stress,the expression of ChAI was predicted to be related to anti-Al stress responsive mechanism,but its accurate function and anti-Al stress mechanism were still elusive and needed to be further investigated.
出处
《核农学报》
CAS
CSCD
北大核心
2010年第6期1286-1290,共5页
Journal of Nuclear Agricultural Sciences
基金
国家自然基金重点项目(30830079)
国家自然科学基金(No.30671419,30700541)
国家高技术研究发展计划863专项经费(2008AA10Z150,2006AA10Z1A8,2006AA100108)
关键词
甜瓜属野生种
表达序列标签
克隆
铝诱导
Cucumis hystrix Chakr.
expressed sequence tags
clone
aluminum-induced
