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

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

用HPLC-MS-MS快速分析和鉴定三尖杉植物内生真菌发酵液中的Brefeldin A

采用HPLC -MS -MS联用技术 ,分析了C56和C65两株具有抗肿瘤活性的三尖杉植物内生真菌发酵液抽提物 ,首次报道了这两株真菌都能产生BrefeldinA(BFA)。采用ESI-MS总离子流跟踪分析HPLC的洗脱液 ,并用低能量的CID -MS -MS(碰撞诱导裂解方式 )进一步确定目标离子峰为BFA分子离子峰 ,这为植物内生真菌发酵液中的有效成分的早期鉴别奠定了基

中国海及邻近区域碳库与通量综合分析

中国海总面积约470万平方公里,纵跨热带、亚热带、温带、北温带等多个气候带.其中,南海北依\"世界第三极\"青藏高原、南邻\"全球气候引擎\"西太平洋暖池,东海拥有全球最宽的陆架之一,跨陆架物质运输显著,黄海是冷暖流交汇区域,渤海则是受人类活动高度影响的内湾浅海.中国海内有长江、黄河、珠江等大河输入,外邻全球两大西边界流之一的黑潮.这些鲜明的特色赋予了中国海碳储库和通量研究的典型代表意义.文章从不同海区（渤海、黄海、东海、南海）、不同界面（陆-海、海-气、水柱-沉积物、边缘海-大洋等）,以及不同生态系统（红树林、盐沼湿地、海草床、海藻养殖、珊瑚礁、水柱生态系统等）多层面对海洋碳库与通量进行了较系统地综合分析,初步估算了各个碳库的储量与不同碳库间的通量.就海气通量而言,渤海向大气中释放CO2约0.22Tg Ca-1,黄海吸收CO2约1.15Tg Ca-1,东海吸收CO2约6.92～23.30Tg Ca-1,南海释放CO2约13.86～33.60Tg Ca-1.如果仅考虑海-气界面的CO2交换,中国海总体上是大气CO2的\"源\",净释放量约6.01～9.33Tg Ca-1.这主要是由于河流输入以及邻近大洋输入所致.河流输入渤黄海、东海、南海的溶解无机碳（DIC）分别为5.04、14.60和40.14Tg Ca-1,而邻近大洋输入DIC更是高达144.81Tg Ca-1,远超中国海向大气释放的碳量.渤海、黄海、东海、南海的沉积有机碳通量分别为2.00、3.60、7.40、7.49Tg Ca-1.东海和南海向邻近大洋输送有机碳通量分别为15.25～36.70和43.39Tg Ca-1.就生态系统而言,中国沿海红树林、盐沼湿地、海草床有机碳埋藏通量为0.36Tg Ca-1,海草床溶解有机碳（DOC）输出通量为0.59Tg Ca-1;中国近海海藻养殖移出碳通量0.68Tg Ca-1,沉积和DOC释放通量分别为0.14和0.82Tg Ca-1.总计,中国海有机碳年输出通量为81.72～103.17Tg Ca-1.中国海的有机碳输出以DOC形式为主,东海向邻近大洋输出的DOC通量约15.00～35.00Tg Ca-1,南海输出约31.39Tg Ca-1.综上,尽管从海-气通量看中国海是大气CO2的\"源\",但考虑了河流、大洋输入、沉积输出以及微型生物碳泵（DOC转化输出）作用后,中国海是重要的储碳区.需要指出的是,文章数据是基于中国海各海区碳循环研究报道,鉴于不同研究方法上的差异,所得数据难免有一定的误差范围,亟待将来统一方法标准下的更多深入研究和分析.国家重点研发计划项目(编号:2016YFA0601400);;国家自然科学基金项目(批准号:91751207、91428308、41722603、41606153、41422603);;中央高校基础研究项目(编号:20720170107);;中海油项目(编号:CNOOC-KJ125FZDXM00TJ001-2014、CNOOCKJ125FZDXM00ZJ001-2014)资

类泛素蛋白及其中文命名

泛素家族包括泛素及类泛素蛋白,约20种成员蛋白.近年来,泛素家族领域取得了迅猛发展,并已与生物学及医学研究的各个领域相互交叉.泛素家族介导的蛋白质降解和细胞自噬机制的发现分别于2004和2016年获得诺贝尔奖.但是,类泛素蛋白并没有统一规范的中文译名. 2018年4月9日在苏州召开的《泛素家族介导的蛋白质降解和细胞自噬》专著的编委会上,部分作者讨论了类泛素蛋白的中文命名问题,并在随后的\"泛素家族、自噬与疾病\"(Ubiquitinfamily,autophagy anddiseases)苏州会议上提出了类泛素蛋白中文翻译草案,此草案在参加该会议的国内学者及海外华人学者间取得了高度共识.冷泉港亚洲\"泛素家族、自噬与疾病\"苏州会议是由美国冷泉港实验室主办、两年一度、面向全球的英文会议.该会议在海内外华人学者中具有广泛影响,因此,参会华人学者的意见具有一定的代表性.本文介绍了10个类别的类泛素蛋白的中文命名,系统总结了它们的结构特点,并比较了参与各种类泛素化修饰的酶和它们的生物学功能.文章由45名从事该领域研究的专家合作撰写,其中包括中国工程院院士1名,相关学者4名,长江学者3名,国家杰出青年科学基金获得者18名和美国知名高校华人教授4名.他们绝大多数是参加编写即将由科学出版社出版的专著《泛素家族介导的蛋白质降解和细胞自噬》的专家

一种聚变反应堆装置的新概念

本文提出了一个聚变反应堆装置的新概念,它由两个轴对称的类似串联磁镜的部分和两个螺旋仿星器U形弯曲段组成,并综合了直管和环形、快过程和慢过程的优点。文中研究了起动、过渡和运行阶段,讨论了D-D反应堆参数和稳定性。射频和高能分量只在起动阶段才需要,在整个过程中只有瞬时的能量消耗。端塞也只须在起动阶段存在,这大大降低了工程技术要求

Progress on Gene Transformation of Cucumber

综述了黄瓜转基因研究所取得的进展,从黄瓜再生体系的建立,遗传转化方法及其影响因素等方面进行了评述,并对黄瓜转基因的不足及今后研究的重点进行了讨论.Cucumber(Cucumis sativus L.) has been extensively investigated for genetic engineering.Cucumber regeneration systems,genetic transformation methods and factors influencing transformation have been reviewed in this paper.Meanwhile,the problems and the research focus of cucumber transformation were discussed.漳州师范学院院内项目(SK05010)资

Expression profiling of key genes involved in taxuyunnanine C biosynthesis in suspension cultures of Taxus chinensis by sucrose feeding and double elicitation with 2,3-dihydroxylpropanyl jasmonate

Taxus suspension cell culture has the potential to provide a sustainable source of anticancer drug paclitaxel (Taxol®) and other taxoids. In the cell culture of Taxus chinensis, taxuyunnanine C (Tc) is the primary taxoid. To design a rational strategy for redirecting the precursor fluxes from other taxoids into paclitaxel production, this study employed quantitative RT-PCR to understand the dynamic profiling of key biosynthetic pathway genes of palcitaxel and taxoids during the cultureprocess. Six genes (TASY,TDAT, T5 H, T H, TDAT and T10 H) were quantified under a process condition of double elicitation by 2,3-dihydroxylpropanyl jasmonate(DHPJA) (100 M) on days 7 and 12, and sucrose feeding (20 g/L) on day 7. This process treatment led to a high accumulation of taxuyunnanine C (Tc) at 554.46±21.28 mg/L 8 days after the first elicitation, 5.5-fold of that of control. And 9 days after the second elicitation, Tc production was as high as 997.72±1.51 mg/L, 9.7-fold of that of control. The early pathway genes TASY and TDAT were signifi- cantly up-regulated by 182-fold and 98-fold, respectively for the first DHPJA elicitation and by 208-fold and 131-fold, respectively for the second elicitation. The induction after each elicitation lasted for about 24 h before their abundances decreased. Things are some different in the case of the other four genes T5 H, T H, TDAT and T10 H. For gene T H, it was greatly up-regulated by 3061-fold for the first DHPJA elicitation and by 1016-fold for the second elicitation. For the other three genes T5 H, T10 H, T14 H, they were up-regulated by 13-fold, 38-fold and 20-fold, respectively for the firstDHPJA elicitation and by 7-fold, 16-fold and 6-fold, respectively for the second elicitation

Studies on the Technical System of AntherCulture of Brassica oleracea var. acephala

本实验以6份羽衣甘蓝为材料进行花药培养.细胞学观察表明,当花蕾长为2.5-3.5mm,花瓣与花药长度之比为1/2~4/5时,45-65%的小孢子发育处于单核晚期,是花药培养的最佳时期.花药培养结果可见基因型不同,胚状体的诱导率有极大的差异,最容易形成胚状体的材料是Y3,诱导效率达17.74%;胚状体的出现在花药接种后的15-30天之间.培养基的种类不同,培养效果也有差异,其中以Keller培养基效果最佳;培养基中有机成份加倍,则可以显著提高胚状体的诱导效率.培养基中蔗糖的浓度对胚状体的诱导率也有显著的影响.Six cultivars of Brassica oleracea var. acephala were used in the study anther culture. The main results are as follows: The cytological observations of pollen showed that most of the pollen were in the late uninucleate stage when the ratio of petal to anther length was between 1/2~4/5 in Brassica oleracea var. acephala ,and the stage of late uninucleate was optimal for anther culture. The effects of genotype and cultural medium on embryoid induction were investigated in s Brassica oleracea var. acephala. The results showed it was optimal to induce the embryoid in Keller medium with double organic ingredients and 10% sucrose. The capacity of embryoid production varied significantly in different genotypes, and genotype Y3 was an ideal material for embryoid induction