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刚竹属几种重要散生竹Lhca1基因的克隆及序列比较.doc

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刚竹属3种重要散生竹光系统Ⅰ基因(Lhca1) 的克隆、序列分析和蛋白结构预测 唐文莉1彭镇华1 2高健1* (1 国际竹藤网络中心 国家林业局竹藤科学与技术重点开放实验室 2 中国林业科学研究院林业研究所) 摘要:光系统在植物光合系统中具有重要功能,Lhca1是编码PS复合物中最主要的捕光色素蛋白LHC的基因。本研究采用RT-PCR法,从毛竹中克隆了捕光叶绿素a/b结合蛋白基因LhcaPe01的全长(GenBank EU035496),从红壳竹和角竹中克隆了长度分别为616bp、613bp的捕光叶绿素a/b结合蛋白基因片段LhcaH01(GenBank EU513200、 LhcaJ01(GenBank EU513201)。运用生物信息学方法对其核苷酸序列、编码的氨基酸序列以及蛋白结构进行预测。结果表明LhcaPe01基因序列从第39bp开始到第779bp含有1个开放阅读框(open reading frame, ORF)和一个中止密码子,该基因全长1051bp,在5’端有38bp的非编码区(untranslated region, UTR),在3’端含有242bp的非编码区(untranslated region, UTR)和Poly(A) 30bp。对这3个基因的保守片段的序列分析表明,刚竹属三种竹种毛竹、红壳竹和角竹保守区内核酸序列同源性非常高,红壳竹与毛竹和角竹的同源性分别为98.1%和98.9%,而毛竹和角竹的同源性高达99.7%;在禾本科内不同属之间毛竹与大麦序列同源性最高,次之,同源性分别为;编码的氨基酸序列同源性与核酸序列同源性一致,最高是毛竹与大麦,水稻次之。经推测LhcaPe01编码的蛋白质等电点和分子量分别为5.4000和22107.34Da。蛋白质结构预测表明,毛竹、大麦、水稻、玉米的LHC蛋白结构非常相似。这是我国首次报道的竹子光合系统Ⅰ的捕光叶绿素a/b结合蛋白基因的全序列。 (Phyllostachys edulis);(Phyllostachys iridescens);(Phyllostachys fimbriligula);Lhca1; Tang Wen-li1; Peng Zhenhua1,2; Gaojian1*. Cloning and sequence analysis of Lhca1 gene the light harvesting complex from three kinds of Phyllostachys and structure prediction of the protein. 1.Key Labratory of Bamboo and Rattan Science Technology of State Forestry Administration, International Centre for Bamboo and Rattan, Beijing, 100102, P. R. China; 2.Research Institute of ForestryChinese Academy of Forestry ,Beijing, 100091, P. R. China. Light harvesting complexⅠ(LhcI), is a chlorophyll a/b-binding protein, which plays a key role in plant photosynthesis. A full-length cDNA encoding Lhca1 gene was cloned from the first strand of Phyllostachys edulis cDNA through RT-PCR method, named as LhcaPe01 (GenBank EU035496). 616 bp cDNA fragment was cloned from the first strand of P. iridescens cDNA, named as LhcaH01 (GenBank EU513200) and 613 bp cDNA fragment was cloned from the first strand of P. fimbriligula cDNA, named as LhcaJ01 (GenBank EU513201). The length of LhcaPe01 is 1051bp,which contains an open reading frame encoding 249 amino acids from 39 to 779 positions and has 38bp an
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