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Cloning and differential expression analysis of a new rbcS gene from Lemna gibba

机译:Lemna gibba新的rbcS基因的克隆和差异表达分析

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摘要

A novel Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) small subunit gene (named ssu4d) was cloned from Lemna gibba by a novel chromosome walking technology. The full-length of ssu4dcDNA (named ssu4dc), contained a 522 bp open reading frame encoding a protein of 174 amino acids. Sequence analysis of ssu4dc and ssu4d showed that ssu4d contained an intron between +355 nt to +1125 nt downstream of transcriptional iniative site. ssu4dc contained 54 bp of 5' untranslated region (UTR), and an open reading frame of 174 amino acids consisting of a chloroplast transit peptide with 57 amino acids and a mature protein of 117 amino acids. The deduced amino acid sequence of ssu4dc shared 95-96% identity with L. gibba RbcS protein. Real time-PCR analysis showed differential expression of individual rbcS genes in light-grown Lemna gibba. And the levels of SSU4dc mRNA was regulated by the action of phytochrome, there was variability in the amount of expression of SSU4dc RNA comprared to the SSU1 and SSU5B from Lemna gibba.
机译:利用新的染色体步移技术从Lemna gibba克隆了一个新的Ribulose-1,5-bis磷酸双羧化酶/加氧酶(Rubisco)小亚基基因(命名为ssu4d)。 ssu4dcDNA的全长(命名为ssu4dc)包含一个522 bp的开放阅读框,编码174个氨基酸的蛋白质。 ssu4dc和ssu4d的序列分析表明,ssu4d在转录起始位点下游+355 nt至+1125 nt之间含有一个内含子。 ssu4dc包含54 bp的5'非翻译区(UTR),以及一个174个氨基酸的开放阅读框,其由具有57个氨基酸的叶绿体转运肽和117个氨基酸的成熟蛋白组成。推导的ssu4dc氨基酸序列与L. gibba RbcS蛋白具有95-96%的同一性。实时荧光定量PCR分析显示,在轻度生长的Lemna gibba中单个rbcS基因的差异表达。 SSU4dc mRNA的表达受植物色素的调节,与Lemna gibba的SSU1和SSU5B相比,SSU4dc RNA的表达量存在差异。

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