Screening and preliminary analysis of an endosperm-specific expression gene in rice(Oryza sativa L.)
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摘要: 从一个水稻启动子捕获突变群体中筛选到一个T0及T1代GUS组织化学检测均表现胚乳特异表达的启动子捕获系w9101,其T0、T1代自交后代标记基因分离规律及Southern blot分析表明它是一个单T-DNA插入的启动子捕获系;通过TAIL-PCR获得其T-DNA插入的侧翼序列,并在NCBI上对其侧翼序列进行比对,发现该捕获系T-DNA插入位点位于一个推测的肽转运子基因(暂命名为Peptide transporter 9101,PTR9101)启动子内;w9101不同发育时期不同组织RT-PCR检测表明,PTR9101为胚乳特异性表达基因;生物信息学分析表明PTR9101蛋白整条肽链表现疏水性,其跨膜区域和CHL1(The Nitrate TransporterAtNRT1.1)一样有12个α-螺旋跨膜区,因此推测PTR9101是一个肽转运子基因。Abstract: A promoter trapping line(accession no.w9101),in which the β-glucuronidase(GUS)reporter gene was endosperm-specific expression in both T0 and T1 generations,was identified from a promoter trapping population in rice(Oryza sativa L.ssp.indica).Genetic analysis and the Southern blot hybridization of the marker gene indicated that w9101 was a trapping line with a copy of T-DNA insertion.The flanking sequence of T-DNA insertion site was isolated by TAIL-PCR in w9101.The BLAST result of the sequence showed that the T-DNA insertion site was located in the promoter sequence of a putative peptide transporter gene in chromosome 6 of rice.It was tentatively named as PTR9101.RT-PCR analysis of the PTR9101 gene revealed that it was specifically expressed in rice endosperm.Further analysis by bioinformatics method indicated that the PTR9101 protein was hydrophobic with 12 α-helix membrane-spanning domain and similar to that of CHL1(The Nitrate Transporter AtNRT1.1)in Arabidopsis.It was thus concluded that PTR9101 was a hydrophobic peptide transporter gene.
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Key words:
- Rice(Oryza sativa L.) /
- endosperm /
- promoter trapping line /
- PTR9101
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