| 摘要: |
| [目的]克隆慈竹木质素生物合成酶4CL基因,研究其序列特征,为今后利用该基因调控慈竹木质素的生物合成研究奠定基础.[方法]参照其他物种4CL基因保守区,设计简并引物,以慈竹幼笋cDNA为模板,克隆到1条750 bp的4CL基因保守区片段.以这条保守区片段为模板,结合5′RACE和3′RACE克隆4CL基因全长序列,并对其进行了生物信息学分析.[结果]成功克隆了慈竹4CL基因全序列,该序列CDS区全长为1 674 bp,编码558个氨基酸;氨基酸序列中存在两大4CL氨基酸保守域(SSGTTGMPKGV和GEICIRG),其中前一个氨基酸保守域的第7个氨基酸位点变为M,与常见的L不同,表明慈竹的4CL基因发生了一定变异.系统进化分析表明,该基因(Na4CL)与水稻的Os4CL4和黑麦草的Lp4CL2具有很高的同源性.[结论]克隆了慈竹木质素合成酶4CL基因全长序列,在GenBank上注册为EU327341,可能与慈竹木质素生物合成有关. |
| 关键词: 慈竹 4CL基因 克隆 生物信息学分析 |
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| 基金项目:四川省应用基础研究基金资助项目(05JY029-101)；四川省教育厅项目(2003031022)；四川省青年基金资助项目(041100)；西南科技大学重点项目(2006031030) |
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| Cloning and bioinformation analysis of 4CL gene in Neosinocalamus affinis |
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| Abstract: |
| 【Objective】 4-coumarate:CoA ligase（4CL） plays an important role in ligin biosythesis pathways.Up to now,there are not reports about the gene coding 4-coumarate:CoA ligase of Neosinocalamus affinis.The coning of 4CL gene of Neosinocalamus a〖JP5〗ff〖JP〗inis would be useful for improving the quality of Neosinocalamus affinis as raw materials for paper making.【Method】 The degenerate primers were designed according to 4CL known function of other plant.The conserved sequence of 4CL gene in Neosinocalamus affiniswas cloned using cDNA of the young shoot of Neosinocalamus affinis and degenerate primers as a template and primers respectively.Then,the complete 4CL gene sequence was cloned by 5′RACE and 3′RACE using the conserved cDNA sequence cloned as a template.【Result】 The full-length sequence of 4CL in Neosinocalamus affinis(Na4CL) was successfully cloned in this study.The bioinformational analysis showed that the CDS region of the nucleotide sequence was 1 674 bp,coding 558 predicted amino acids with the conserved amino acid blocks SSGTTGMPKGV and GEICIRG.An active M residue embedded in the SSGTTGMPKGV motif of Na4CL,which is different from L residue in SSGTTGLPKGV motif of 4CL of other plants.Phylogenetic analysis showed that Na4CL was most similar to Os4CL4 in rice and Lp4CL2 in Lolium perenne.【Conclusion】 The 4CL gene named Na4CL was first cloned in Neosinocalamus affinis,which would play an important role in ligin biosythesis pathways of Neosinocalamus affinis.And then the CDS sequence of Na4CL has been submitted to GenBank with the accession number EU327341. |
| Key words: Neosinocalamus affinis 4CL gene cloning bioinformation analysis |
