Expression and Construction of Yeast Expression Vector of LBM2eHBc+ gene in P.pastoris

禽流感病毒一直是家禽和家畜健康养殖的巨大威胁,之前的研究表明,原核系统中表达的lTb和M2EHbC+融合基因的产物能有效诱导免疫动物对禽流感病毒M2E多肽产生特异性的粘膜免疫应答.酵母作为生物反应器应用于生物制品生产具有独特的优点.本研究构建了PPIC9k-lbM2EHbC+酵母表达载体并对酵母gS115进行了遗传转化,甲醇诱导表达结果表明lbM2EHbC+融合基因在酵母细胞中得到表达,表达出的融合蛋白能够被M2抗体识别.Avian influenza virus(AIV) has been a constant threat to the healthy development of livestock and poultry breeding industry.Previous research has shown that the products of LTB and M2eHBc+ fused gene expressed in prokaryotic cells can effectively induce mice mucosal immune responses against M2e epitope.P.pastoris yeast has a unique advantage as a bioreactor used in the production of biological products.LBM2eHBc+ fused gene fragment was obtained by PCR,then inserted into pPIC9K plasmid which contains methanol promoter and yeast signal peptide to construct yeast expression vector pPIC9K-LBM2eHBc+.The recombinant vector was transformed into P.pastoris GS115.The methanol induction result has indicated that the LBM2eHBc+ fusion gene can be efficiently expressed in GS115 cell and the expressing protein has the specific binding activity to M2 antibody by Western blotting analysis..福建省科技厅重大项目子课题(2006NZ0003-2);福建省教育厅A类科技项目(JA09168

Studies on Construction of Plant Expression Vector of Recombinant LBM2eHBc+ Gene and Tomato Transformation

禽流感病毒一直是家禽和家畜健康养殖的巨大威胁,甚至威胁着人类的健康,人们也一直在研究各种形式的疫苗来应对禽流感病毒的威胁.番茄作为生物反应器应用于动物疫苗生产具有独特的优点,本研究优化了番茄子叶的再生体系,同时构建了lbM2EHbC+融合基因的植物表达载体并对番茄进行了遗传转化.结果表明在玉米素(zT)浓度为0.5 Mg/l时番茄子叶外植体的芽再生率达到93.33%;测序结果证明植物表达载体PbI121-lbM2EHbC+构建成功;利用农杆菌介导的转化方法获得再生抗性苗34株,经PCr检测,阳性率为75.0%,本研究结果为进一步开发禽流感口服疫苗奠定了基础.Avian influenza virus(AIV) infect almost all wild bird and domestic poultries and then decimate poultries,even the threat of AIV overhangs human.It also has been researching a variety of vaccines to combat the AIV threat.Tomato as a bioreactor used in animal vaccine production has a unique advantage.This study optimized the regeneration system of tomato cotyledon,while construct LBM2eHBc + gene plant expression vector,and then carried out the genetic transformation of tomato.The results show that the bud regeneration rates of tomato reach the highest 93.33% when MS medium supplemented 0.5 mg/L Zt+ 1 mg/L IAA;sequence results show that plant expression vector pBI121-LBM2eHBc + constructed successfully.A total of 34 putative transgenic tomato plants were obtained by using of Agrobacterium-mediated transformation method,and the positive rate was 75.0% by PCR detection.The results laid a foundation for the further development of oral AIV vaccines.福建省科技厅重大项目子课题(2006NZ0003-2);福建省教育厅A类科技项目(JA09168

Annual change of outdoor seawater cultivation of dunaliella for biomass energy

作者简介：陈昱（1982-），男，硕士，主要从事生物质能源研究。E-mail：[email protected][中文文摘]将涨潮海水浓缩后以次氯酸钠进行消毒,添加少许氮、磷等无机盐,在室外周年养殖杜氏藻及其多个突变株,研究其在自然条件下生物量的产出和细胞内脂类的积累。结果表明,在养殖过程中,培养液的盐度、pH值以及细菌含量的变化对杜氏藻的生长没有显著影响。通过培养,不同株系杜氏藻都可以获得较高的生物量和脂类产出,特别是耐高温突变株、高脂突变株可以适应夏天室外的高温、高盐等恶劣环境,普通株在冬天尚能生长,不同特性的藻株的轮换养殖可以保证养殖场周年养殖。海水养殖有望成为一种大规模培养杜氏藻的既简单又经济的发展模式。文章为进一步开发利用杜氏藻生物质能源奠定了一定的基础。[英文文摘]Dunaliella bardawil and several mutants were cultured outdoor in concentrated seawater treated by Javel water with the addition of few nitrate and phosphate,and the biomass and lipid accumulation of Dunaliella were assessed.Dunaliella species grew well during the cultivation period,despite of the change of salinity,pH and bacteria content in seawater.High lipid and biomass productivity of Dunaliella species was obtained in our study.High temperature tolerance mutant and high lipid content mutant grew well under high temperature and high salinity of seawater in summer, while normal strain grew well in winter. Alternate cultivation of different Dunaliella strain should be carried out in all-year outdoor cultivation. It would be a simple, economic batch mode for large-scale Dunaliella cultivation using concentrated seawater, and it would contribute to further using of Dunaliella for biomass energy.福建省科技重点项目(2009N0052); 福建省重大专项前期研究项目(2005YZ1022

Brown tide:A new ecosystem disruptive algal bloom

AurEOCOCCuS AnOPHAgEffErEnS和AurEOuMbrA lAgunEnSIS隶属于棕鞭藻门(OCHrOPHyTA)、海金藻纲(PElAgOPHyCEAE),在美国及南非的一些河口形成生态系统破坏性褐潮(brOWn TIdE)已经有20多年了,近年来在中国河北沿海发生的大规模褐潮也使中国成为世界上第3个受褐潮影响的国家。褐潮藻能够利用多种有机营养,在低光照及低营养条件下达到高生长速率,对贝类养殖产业、经济以及娱乐产业等造成严重的负面影响。本文总结了20多年来褐潮在全球的发生及其所造成的严重危害,对褐潮藻的形态结构、生理特征及分子遗传学,特别是促进褐潮形成及持续的关键蛋白的编码基因等研究作了简要介绍,并概括了国内褐潮的研究现状。在此基础上,展望今后褐潮的研究方向,以期为褐潮的研究、预警预报和生态学防治及其防灾减灾提供借鉴。The pelagophytes Aureococcus anophagefferens and Aureoumbra lagunensis have formed ecosystem disruptive algal blooms in shallow estuaries of the United States and South Africa for more than two decades.The large-scale brown tide events in the coastal waters of Hebei Province in recent years make China the third country that is affected by brown tide in the world.These algae are able to utilize a wide variety of organic nutrients and achieve high growth rates at low light and nutrient levels.They have significant negative impacts on the shellfish mariculture industry,economy and recreational industry.We review research progress in the last two decades on morphology,physiology and molecular and genomic characteristics of these brown tide algae,especially the genes that encode many of the key proteins that facilitate bloom formation and persistence.Moreover,the research progress in China about brown tide is summarized.The prospects of the research on brown tide are also discussed.海洋公益性行业科研专项(201205031-03和201005015-5); 国家基础研究发展计划项目(2010CB428704)资

The Effects of Different Lights and Gibberellin on Establishment of Parasitism between Dodder and Its Hosts

首次使用lEd作为光源,研究不同光照条件及gA3对菟丝子(CuSCuTA SPP.)弯钩打开、缠绕发生与吸器形成的影响。结果表明,光照信号作为一个必要条件参与了菟丝子对寄主的识别及缠绕发生的调控,而化学信号可能起到一定的促进作用;gA3参与了对菟丝子缠绕发生的调控,但对弯钩打开没有明显的作用。除了典型的光敏色素作用外,还有另一类光反应(HEr)参与了上述过程,这类光反应可由879 nM远红光引发,证明菟丝子存在HEr,还有Pfr向Pr的暗转化过程,在缠绕发生过程中光敏色素和隐花色素发生相互作用。The effects of different light treatments and GA3 on hook opening,twining and parasitism of Cuscuta australis were studied using LED as light sources for the first time.The results showed that light was a necessary factor for dodders to parasitize the hosts successfully and chemical signals might facilitate host recognition and twining.GA3 involved in controlling twining response,but no distinct effect on hook opening.Furthermore,besides typical(phytochrome reaction,another photoreaction called HER was involved in these processes,which could be caused by 879 nm far red light.So it demonstrated directly that there was not only HER in dodders,but also dark conversion from Pfr to Pr,and there were mutual interaction of phytochromes and cryptochromes in twining.教育部重点项目(101102)资

Study on Tomato Transformation with Human Keratinocyte Growth Factor-1

人角质细胞生长因子在组织的损伤修复中起着重要的作用.以番茄子叶为外植体,通过根癌农杆菌介导法,将人kgf-1导入番茄中,共获得12株独立的抗性转化植株,PCr和dnA斑点杂交结果表明:kgf-1基因整合入番茄基因组中,为获得植物源表达的kgf-1蛋白打下了基础.Keratinocyte growth factor plays an important role in the tissue damage repair.Using sterile tomato cotyledons as explants,the human KGF-1 gene had been introduced into the tomato genome by Agrobacterium-mediated transformation method.As a result,a total of 12 independent putative transgenic tomato plantlets were obtained.The results of PCR and DNA dot-blot hybridization showed that KGF-1 gene was integrated into the tomato genome.This work has laid a foundation for further studies on the development of KGF-1 proteins expressed in tomatoes.福建省自然科学基金(2010J01240

SX微处理器及其在数字电感测试仪中的应用

SX微处理器是美国SCENIX公司推出的新一代 8位微处理器 ,本文介绍了该微处理器的功能和特点 ,并给出了它在数字电感测试仪中的应用实例

Expression of Modified H5N1 Avian Influenza Virus M2 Gene and the Recombinant M2 Protein Immunogenicity Assay

禽流感的发生不仅会造成禽类的大量死亡,而且也严重地威胁着人类健康,接种疫苗是控制禽流感发生的主要措施之一.M2基因的保守性使其成为基因工程亚单位疫苗的目标抗原.本研究将M2基因的跨膜区删除后,构建原核表达载体PET32A-△M 2,IPTg诱导后,收获的细菌总蛋白SdS-PAgE电泳结果表明修饰的M2基因在原核表达系统中得到高效表达,表达蛋白以可溶性形式存在,WESTErn杂交和动物免疫结果表明重组蛋白具有抗原性和免疫原性.本研究结果为利用M2重组蛋白开发具有交叉保护作用的禽流感疫苗奠定了基础.The present vaccination is one of main strategy for control of avian influenza virus that critically threatened human health.Recent investigations of subunit component vaccine focused on M2 gene because M2 gene possessed a highly structurally conserved property among influenza viral strain.In this study,the recombinants prokaryotic expression plasmid pET32a-△ M2 harboring the deletion of the transmembrane segment of M2 gene was constructed.SDS-PAGE electrophoresis showed that modified M2 gene was highly expressed as fusion protein in a soluble form after genetic modified E.coli BL21(DE3) bacteria were induced with IPTG.The fusion protein of M2 possessed antigenicity and immunogenicity properties by Western Blotting analysis of fusion protein with standard antibody against M2(H5N1) and mouse vaccination of purified fusion protein respectively.The results laid a foundation of further study on the development of cross-protection avian influenza vaccine.福建省科技厅重点项目(2009N0051);福建省教育厅A类科技项目(JA09168

Compass三维剂量验证系统在鼻咽癌容积旋转调强放射治疗计划剂量验证中的应用

目的:探讨Compass三维剂量验证系统在鼻咽癌容积旋转调强放射治疗(VMAT)计划剂量验证中产生的剂量差异,并分析其原因。方法:选取17例鼻咽癌患者,在Monaco上制定VMAT计划并将其传至Compass,运用Compass自身计算的剂量(CD)和通过实测重建的剂量(RD)两种方法验证靶区和危及器官(OAR)的γ通过率、D1%、D99%、Dmean等参数。结果:治疗计划系统(TPS)-CD的所有γ通过率均>97.5%,TPS-RD的所有γ通过率均>95.0%,且TPS-CD的每一个γ通过率均大于TPS-RD中对应的γ通过率,两者靶区的γ通过率比较差异有统计学意义(t=2.110,t=2.749,t=2.489,t=2.687,t=2.798,t=2.881,t=2.921;P<0.05),但两者在OAR比较则无统计学意义。TPS-RD的靶区D1%、Dmean及D99%的三项指标绝对剂量差均<200 cGy,百分差<2.5%,且这三项指标在TPS和RD之间的差异均无统计学意义;左右晶状体的三项指标的剂量在TPS和RD之间的差异均有明显统计学意义(t=4.328,t=4.658,t=4.210,t=4.511,t=4.896,t=5.241;P<0.05);脊髓、脑干、左右腮腺等OAR的D99%在TPS和RD之间的差异有统计学意义(t=4.018,t=4.035,t=3.646,t=4.112;P<0.05),Dmean和D1%在TPS和RD之间的差异均无统计学意义。结论:Compass三维剂量验证系统在VMAT计划剂量验证中可直观、快速的分析出靶区和OAR的理论和实际照射剂量差异,为剂量的精准实施提供数据支持

Cloning and Characterization of Na~+/H~+ Antiporter Gene (nhaA) from Pseudomonas sp.cn4902

根据3种生物的Na+/H+逆向转运蛋白基因(nhaA)的两端序列设计引物,利用PCR从假单胞菌 (Pseudomonassp.cn4902)中克隆得到一结构基因。该基因长1089bp,编码362个氨基酸,与E.coliK12的 nhaA基因的同源性高达97.0%。将该结构基因与pBV220构建成重组载体pBVA。SDS PAGE电泳表明:含 pBVA的转化子产生较高浓度的分子量约为41kD的蛋白,与预期相符。在含NaCl1.0mol/L的培养基中生长达 到平衡期时,转化子的菌浓度约是对照的2.3倍。经原子吸收光谱测定,转化子细胞质中Na+浓度仅为对照菌的 60.4%。SDS PAGE电泳表明该基因的表达蛋白位于细胞膜(壁)上。提纯外源基因表达蛋白并对其N端8个氨 基酸进行测序,与nhaA基因推测的氨基酸序列完全相符。这些实验证实,克隆得到的基因是假单胞菌的nhaA基 因。该基因已经在GenBank登记,收录号为AY643494。 【英文摘要】 According to the sequences of the gene nhaA coding for Na~+/H~+ antiporter,a structural gene was cloned from Pseudomonas sp.cn4902 by PCR reaction with a set of primers.It was 1 089 bp in length and codes for 362 amino acids sharing homology with the gene nhaA of E.coli K12 as high as 97.0%.It was inserted into plasmid pBV220 to form a high level expression reconstruction plasmid pBVA.So an overexpression 41 kD protein band could be found in the lane of transformant harbored with pBVA after SDS-PAGE electro...福建省科技计划重点资助项目(编号:2003N053);; 厦门大学生命科学学院细胞生物学与肿瘤细胞工程教育部重点实验室项目(编号:2004106)~