Cloning and Sequence Analysis on Gene of Chlorophyll a/b-binding Protein from Narcissustazetta var. chinensis
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摘要: 以多效唑诱导中国水仙的抑制差减杂交文库(SSH)获得的cDNA片段为基础,采用RACE及RT-PCR技术,分离克隆到1个叶绿素a/b结合蛋白基因Ntcab1。此基因包含有1个798bp完整阅读框架(ORF),编码265个氨基酸,分子量为28.155kD,理论等电点为5.48。聚类分析表明:其氨基酸序列与拟南芥LHCb1.1聚为一类,推测其属于LHCb1类蛋白,与麦冬和石蒜的Cab亲缘关系最近。半定量RT-PCR分析结果表明:喷雾多效唑后,Ntcab1在抽葶期和始花期的表达量比喷雾清水稍高。
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关键词:
- 中国水仙 /
- 叶绿素a/b结合蛋白 /
- 基因克隆
Abstract: From a cDNA fragment of the suppression subtractive hybridization of Narcissus tazetta var.chinensis induced by paclobutrazol,the gene of the light-harvesting chlorophyll a/b-binding protein,Ntcab1,was isolated and cloned by RACE and RT-PCR.The full length of Ntcab1cDNA was determined to be 1 012bp with a 798bp ORF,which encoded a protein of 265 amino acids with a predicted molecular weight of 28.155kD and a theoretical isoelectric point of 5.48.The phylogenetic analysis showed that Ntcab1could be clustered together with LHCb1.1 from Arabidopsis thaliana,and had a close relationship with those in Ophiopogon japonicus(ACD64978.1)and Lycoris radiata(ADC35579.1).The semi-quantitative RT-PCR revealed that the expression of Ntcab1was higher during sprouting and early flowering after it was sprayed with paclobutrazol than water. -
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