Cloning and prokaryotic expression of the main subunit allophycocyanin genes from Porphyra haitanensis

别藻蓝蛋白是一种藻胆蛋白，作为捕光和激发能传递物质，在藻类的光合作用中起重要的作用；作为储存蛋白，是一种价值很高的营养成分。不仅如此，藻胆蛋白在免疫荧光技术、抗肿瘤药物开发、医疗保健药物开发、食品工业、精细化工和化妆品工业等许多方面有很重要的应用价值和研究前景。别藻蓝蛋白主要由α、β两亚基聚集构成，每个亚基上均含有一个藻蓝胆素分子。别藻蓝蛋白α、β两亚基分别由叶绿体基因组编码，并且两亚基基因相毗连，中间只有少数碱基的间隔序列。 本课题以阴干的坛紫菜叶状体为材料，通过酶解等方法获取完整的叶绿体，再裂解叶绿体并酚仿抽提获得叶绿体DNA。限制性内切酶HindIII和EcoRI双酶切及RAPD检测结...Allophycocyanins, one of phycobiliproteins, acted as substances of light-havesting and transferring energy to photosystem reaction centers in algal photosynthesis；also as a kind of valuable nutritious components as store proteins；moreover,many researches revealed that allophycocya- nins had extensive applied perspective in quite a lot of fields,such as immunofluorescence technique, anti-tumour med...学位：理学硕士院系专业：生命科学学院生物学系_动物学学号：20002601

A Method of Rapidly Extracting Chloroplast DNA from Porphyra haitanensis

以阴干的坛紫菜叶状体为材料,通过酶解等方法获取完整的叶绿体,再裂解叶绿体并酚仿抽提获得叶绿体DNA.限制性内切酶HindⅢ和EcoRⅠ双酶切及RAPD检测结果显示:叶绿体DNA与基因组DNA的酶切图谱及RAPD图谱均有比较明显的差异.多次重复实验证明,按此法提取的叶绿体DNA,其得率稳定,并可用作PCR扩增的模板及酶切图谱的构建等.该方法操作简便,过程快捷,可作为大型海藻叶绿体DNA提取的参考方法.A convenient method for rapid isolation of chloroplast DNA from Porphyra haitanensis was presented. First, the chloroplasts were isolated, and then lysed in sarcosylproteinase K solution, after that the chloroplast DNA was extracted and purified by phenolchloroform method. The result showed that the chloroplast DNA had different bands from the genomic DNA when double digested with restriction enzyme EcoRⅠ and HindⅢ or RAPD tested. The quality and quantity of the obtained chloroplast DNA was steady and credible by several repeated experiments. It could be used as the template of PCR and construction of physical map. Even more, this method could be the reference method when preparing other algal chloroplast DNA.科技部海洋863项目(2002A603023);; 福建省重大农业科技项目(2001Z017)资

Cloning and sequence analysis of allophycocyanin genes from Porphyra haitanensis

以野生坛紫菜色素体DNA为模板, 通过PCR扩增获得编码坛紫菜别藻蓝蛋白α亚基和β亚基的序列及两者之间的间隔序列,该序列全长为1 055 bp,其中编码α和β亚基的序列均为486 bp,间隔序列为83 bp.该序列与GenBank收录的其他4种红藻相关序列的同源性在75.6%~87.4%,与2种蓝藻的同源性分别为66.9%,68.5%,其中编码区同源性更高.On the basis of the sequences of allophycocyanins from several species in genbank,a pair of degenerate primers were designed and synthesized. A specific fragment about 1 055 bp in size was obtained after PCR amplification using the cpDNA of Porphyra haitanensis as template. Then the purified fragment of DNA was cloned into vector pMD18-T, and sequenced after the identification of the recombinants using PCR and endonuclease digest. The results indicate that α and β subunit genes of allophycocyanin are obtained from Porphyra haitanensis. The fragment size is (1 055 bp), including α and β subunits coding regions (486 bp) each and the spacer between them is 83 bp.The genes were submitted to genbank and their genbank accession number was AY372218.福建省重大农业科技资助项目(2001Z017);; 国家海洋“863”资助项目(2002AA603023

展望分子海洋药物学

随着现代生物技术的发展,海洋药物的研究已达到分子水平。本文从海洋药用生物的鉴定及其分类进化、分子遗传标记育种、外源药物基因表达、病虫害防治、药用有效成分的代谢调控、海洋生物药物功能基因的研究和开发等六个方面对分子海洋药物学进行了概述和展望

藻类基因工程的一些进展

<正> 一、藻类基因工程的发展前沿是基因组学研究1990年启动的人类基因组计划已于2003年4月完成。这个计划除了完成人类染色体DNA中约30亿对碱基排列顺序的测定外,还带动了一批模式生物基因组DNA序列的测定。这些结构基因组学研究成果与以后进行的功能基因组学研究进展,将使我们对生命活动的理解深入到基因水平,对生物物种的改良和对所需生物产品的生产提高到空前的水平。从1996年完成了集胞藻6803的基因组全序列测定以后,到2003年底,已有10种原核和真核藻的基因组的DNA序列完成了测定(表1)