Studies on Transformation of Tomato with Vaccine of Recombinant M2 Peptide of AIV and Keratinocyte Growth Factor

禽流感(AvianInfluenza,AI)是由正粘病毒科A型流感病毒引起的一种以侵害呼吸系统为主的高度接触性传染病。其高致病力毒株可引起90%以上的死亡率，每次爆发都给养禽业带来巨大的经济损失，因此被国际兽疫局列为Ⅰ类烈性传染病。禽流感病毒M2蛋白是由97个氨基酸组成的寡四聚体，与HA和NA基因相比，M2基因具有高度保守性。因此，与其他禽流感多肽疫苗相比，M2蛋白具有交叉保护能力的“通用”流感疫苗研究的候选抗原的优势。 人角质细胞生长因子(Keratinocytegrowthfactor,KGF)是由163个氨基酸残基组成的26-28KD单链多肽。KGF对皮肤创面的上皮细胞具有促进增殖、促...Avian influenza virus (AIV) infect almost all wild birds and domestic poultries and then decimate poultries. Severe economic loss of the poultry industry was brought because of the AIV prevalence. The M2 protein of AIV is a kind of cross-protection constructed by 97 amino acids , and a high evolutionary conservative protein in different subtypes. Because AIV infects animals by the respiratory and ...学位：理学硕士院系专业：生命科学学院生物学系_发育生物学学号：2012005130204

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

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

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

An LVMM Algorithm for the Identification of Arabidopsis Polyadenylation Sites and SNPs-associated Study

真核生物的基因表达需要经过转录和翻译两个过程。在转录过程中，成熟mRNA要经过pre-mRNA的内含子剪接、5’端加帽和3’末端的多聚腺苷化加工（简称poly(A)位点）才能形成。其中，多聚腺苷化能标记出基因的终止位置，决定着基因结构，影响mRNA的稳定性，且在生物基因组中大量存在的单核苷酸多态性（SingleNucleotidePolymorphism，SNP）是生物个体间表现出多态性的原因之一。研究表明，若基因中含有多个poly(A)位点时，位于基因内各区域（如3’UTR）的SNPs可能会通过改变poly(A)信号或位点上下游各碱基含量影响mRNA的3’末端poly(A)位点选择，从而引起...In eukaryotic organisms, precursor messenger RNA(pre-mRNA), after being transcribed from its coding gene, has to undergo processing events such as 5’ capping, intron splicing and 3’ end polyadenylation(poly(A)), then becomes mature and functional mRNA during the transcription process. Therein, the 3’ end polyadenylation processing not only can mark the end of a gene, thus the structure of a gene, ...学位：工学硕士院系专业：信息科学与技术学院_系统工程学号：2322010115326

高炉炉底温度分布

本文提供了一种计算高炉炉底温度分布的方法,这对炉底设计和衬里材料的选择是很有意义的。由于铁液渗透,炉底渗透区材料的导热系数发生变化,铁液固相线(1150℃)会向下移动,我们应用迭代技术,确定固相线的可能分布区域及其极限位置。二维稳定温度场采用有限元素法作数值求解。本文对不同耐火材质的炉衬和有、无底部冷却的不同条件进行了计算分析。初步认为,对大型高炉,底部有冷却的全碳砖炉底结构是比较好的