【作者】 任琛；

【导师】 李名扬；

【作者基本信息】 西南大学 ， 细胞生物学， 2006， 硕士

【摘要】 蜡梅学名：Chimonanthus praecox(L．)Link，英文名：Wintersweet，科名：蜡梅科Calycanthaceae。蜡梅为落叶灌木，原产于我国江苏、浙江、安徽、湖北、江西和陕西等地。在我国栽培己久，是有名的色香俱佳的冬季花卉，花期长达3个月之久。花可提取芳香油、并为解毒生津药，花蕾油治烫伤，根、茎能作镇咳、止喘药。然而作为一种名贵花卉，对蜡梅的分子生物学研究却并不多见。 在分子生物学技术不断取得前进的前提下，人们发现在植物的花药发育和花粉成熟过程中，有大量的基因表达，其中有许多是花药和花粉组织特异性的，目前花药和花粉特异性表达的基因及其调控序列已经成为研究的热点。 本实验的研究内容主要是：从本试验室已构建的蜡梅cDNA文库中分离克隆了pollen specific protein基因，对其进行了分子特性分析以及对其可能的功能进行功能预测，并且构建了该pollen specific protein基因的表达载体，其主要研究如下： 1．克隆子的获得 从本实验室已建完成的cDNA文库中随机挑选文库克隆，提取质粒DNA，采用ABI3700 DNA Sequence DNA自动序列测定仪进行正向序列测定得到EST序列(pspressed sequence tags，ESTs)DW223488。根据EST，以文库克隆载体pTriplEx2引物扩增出约900bp的特异片段，推断其为EST序列所对应的全长cDNA。 2．序列特征分析对目标基因在NCBI(http://www.ncbi.nlm.nih.gov/)上比对分析。发现该基因由815个核苷酸组成，包含一个480个核苷酸的ORF，编码蛋白长159个氨基酸。该蛋白理论分子量为17.5kD，预测等电点为6.24。BlastN分析发现和拟南芥的SAH7基因最相似；BlastP比对发现目标克隆子的EST序列与100条蛋白质序列存在不同程度的相似性，其中排在第一位的与核苷酸比对的结果接近，是葡萄中的花粉特异蛋白pollen-specific protein[Vitis pseudoreticulata](ABC86745)；BlastX分析，它与40个花粉特异蛋白有较高的同源性，其中与葡萄中鉴定的花粉特异蛋白同源性最高，更多还原

【Abstract】 The Chimonanthus praecox. is the fallen leaf bush, originally it produces in places such as Jiangsu, Zhejiang, Anhui, Hubei, Jiangxi and Shanxi.It has been cultivated for a long time in our country.As a famous good winter flowers, the Chimonanthus praecox possesses beautiful colour and pleasant fragrance, and besides ,The flowering season of the Chimonanthus praecox could last for three months long. The flower may withdraw the essential oil, and be a medicine for the disintoxicating and cold.The flower bud oil can cur scalds, the root and the stem can make the town to cough, to stop breathes heavily.However takes one kind of precious plants,not actually have the reseach for the chimonanthus praecox in molecular biology.Under the premise of the advance in the molecular biology technology, people has discovered that in the anther growth and the pollen mature process, has the massive genes expressed. Many of them has been in the anther and the pollen organization specificity, And along with time passing, the disquisition of the anther and the pollen specificity expression gene and its the regulative sequence already becomes the research the hot spot.These are studied on that—Separated the pollen specific protein gene from the chimonanthus praecox cDNA library which has been constructed by our laboratory.And has carried on the molecular characteristic analysis to it as well as to carry on the function to its possible function to forecast, And constructed this pollen specific the protein gene to express the carrier.Its mainly research as follows.1.Get Cppspl gene From cDNA librara which has constructed completes stochastic choice library clone, then distill plasmid DNA , and then, use ABI3700 DNA sequence the DNA —更多还原