木薯果胶甲基酯酶抑制因子MePMEI1的生物信息学分析

为探讨木薯MePMEI1的分子结构特征。通过PCR扩增和测序技术及生物信息学分析工具对木薯MePMEI1基因进行克隆、测序及相关生物信息学分析。结果表明木薯MePMEI1基因编码区全长609 bp,编码202个氨基酸残基;MePMEI1基因编码蛋白分子量21.78 k D,理论等电点(pI)约为5.51;生物信息学预测发现,木薯MePMEI1蛋白是稳定的亲水蛋白;具有跨膜区为分泌蛋白;含有1个PMEI结构域,1个糖基化位点,31个磷酸化位点;二、三级结构以α螺旋和无规则卷曲为主。该蛋白的生物功能可能与细胞被膜、酶和生长因子等相关。木薯MePMEI1基因的生物信息学分析为进一步研究其遗传特性和生理生化机制提供了理论依据。     

幽门螺杆菌HspA融合蛋白口服疫苗的构建

构建表达幽门螺杆菌的保护性抗原分热休克蛋白A亚单位(HspA）和霍乱毒素B亚单位（CtxB）的重组融合蛋白的生物工程菌株,以此制备幽门螺杆菌的口服疫苗。用PCR方法扩增hspA和ctxB两个目的的基因片段,将它们分别克隆至pSK（＋）质粒上,然后插入含T7启动子ET－22b（＋）的表达载体中,构建嗓基因的表达质量pET－hct,转化E.coliBL21(DE3）,经IPTG诱导表达融合蛋白HCT。经测序,hspA-ctxB(hct)融合基因片段由726bp组成,可以编码242个氨基酸残基的多肽。经SDS－PAGE和免疫印迹分析检测发现,融合基因表达的蛋白质相对分子质量约为30kD。融合蛋白经镍离子柱纯化、复性后,和HspA共同标记同位素^125I,然后给小鼠灌胃,结果观察到HCT组小鼠血清中的^125I的放射量要明显高于HspA组（P＜0．001）,且吸收峰值时间明显提前。融合蛋白中的CtxB可明显促进小鼠对HspA的吸收,HCT融合蛋白可以作为预防和治疗幽门螺杆菌感染的侯选口服疫苗。     

Nuclear retention of the lncRNA SNHG1 by doxorubicin attenuates hnRNPC–p53 protein interactions

The protein p53 plays a crucial role in the regulation of cellular responses to diverse stresses. Thus, a major priority in cell biology is to define the mechanisms that regulate p53 activity in response to stresses or maintain it at basal levels under normal conditions. Moreover, further investigation is required to establish whether RNA participates in regulating p53's interaction with other proteins. Here, by conducting systematic experiments, we discovered a p53 interactor—hnRNPC—that directly binds to p53, destabilizes it, and prevents its activation under normal conditions. Upon doxorubicin treatment, the lncRNA SNHG1 is retained in the nucleus through its binding with nucleolin and it competes with p53 for hnRNPC binding, which upregulates p53 levels and promotes p53‐dependent apoptosis by impairing hnRNPC regulation of p53 activity. Our results indicate that a balance between lncRNA SNHG1 and hnRNPC regulates p53 activity and p53‐dependent apoptosis upon doxorubicin treatment, and further indicate that a change in lncRNA subcellular localization under specific circumstances is biologically significant.     

三角酵母D—氨基酸氧化酶基因的克隆,测序及表达

利用跨越内含子的ＰＣＲ技术,从三角酵母（Ｔｒｉｇｏｎｏｐｓｉｓｖａｒｉａｂｉｌｉｓ）变种ＦＡ１１０中扩增得到Ｄ氨基酸氧化酶基因（ｄａａｏ）,并通过ＴＡ克隆的方法将其克隆至ｐＧＥＭＴ载体。序列测定结果表明,所得ｄａａｏ基因的５′端内含子已被删除,基因总长度为１０７１ｂｐ,它与Ｔｒｉｇｏｎｏｐｓｉｓｖａｒｉａｂｉｌｉｓ的Ｄ氨基酸氧化酶同源性达９８３％,与Ｆｕｓａｒｉｕｍｓｏｌａｎｉ和Ｒｈｏｄｏｔｏｒｕｌａｇｒａｃｉｌｉｓ的同源性分别是３８９％和３０８％。为提高表达水平,又将此基因转移至高表达载体ｐＥＴ２８ｂ上,在大肠杆菌ＢＬ２１（ＤＥ３）中进行诱导表达。经ＩＰＴＧ诱导,目的蛋白的产生量可占菌体总蛋白量的４６％,分子量约为３８ｋＤ。Ｄ氨基酸氧化酶的活力可达８０２ｕ／Ｌ。     

南京城市公园绿地不同植被类型土壤呼吸的变化

子2007年10月-2008年9月,利用Li-6400便携式光合作用仪配合土壤呼吸气室对南京中山植物园内草坪、疏林和近自然林3种植被类型的土壤呼吸速率的季节变化及其影响因子进行了测定.结果表明:不同植被类型土壤呼吸速率具有明显的季节变化,夏季(8月)较高,近自然林、疏林和草坪类型分别为3.28、4.07和7.58 μmol·m~(-2)·s~(-1),冬季(12月)最低,近自然林、疏林和草坪类型分别为0.82、0.99和1.42 μmol·m~(-2)·s~(-1);不同植被类型的年均土壤呼吸速率有显著差异(P＜0.05),平均土壤呼吸速率大小排序为草坪>疏林>近自然林;不同植被类型的土壤呼吸速率与土壤温度呈显著性指数相关关系,与土壤含水率无显著相关关系;Q_(10)值均随着土层深度的增加而增加;不同植被类型的Q_(10)值存在一定程度的差异,近自然林类型的Q_(10)值大于草坪和疏林类型的Q_(10)值.研究表明,城市如果大量发展草坪可能增加土壤CO_2的排放.

Abstract：

By using Li-6400 portable photosynthetic apparatus connected to soil chamber,the soil respiration rate under three vegetation types(lawn,open woodland,and close-to-nature forest)in Nanjing Zhongshan Botanical Garden was measured from October 2007 to September 2008,with related affecting factors analyzed.The soil respiration rate had obvious seasonal fluctuation,being the highest in summer(August)and the lowest in winter(December).For the close-to-nature forest,open woodland,and lawn,their soil respiration rate in summer was 3.28,4.07,and 7.58μmol·m~(-2)·s~(-1),and that in winter was 0.82,0.99,and 1.42μmol·m~(-2)·s~(-1),respectively.The annual mean soil respiration rate differed significantly with vegetation type(P＜0.05),which was in order of close-to-nature forest＜open woodland＜lawn.The soil respiration rate had significant exponential correlation with soil temperature,but no correlation with soil moisture.The Q_(10) value increased with increasing soil depth,and was larger in close-tonature forest than in open woodland and lawn.Our results indicated that the rapid development of lawn in urban green space could increase the urban soil CO_2 emission.     

拟南芥转录因子GRAS 家族基因群响应渗透和干旱胁迫的初步探索

GRAS家族是一类植物特有的转录调控因子, 已有报道表明该家族基因在植物生长发育和光信号转导过程中具有重要作用。目前在拟南芥(Arabidopsis thaliana)基因组中已鉴定了33个GRAS家族基因。利用功能基因组学和生物信息学手段,通过基因芯片数据挖掘和基因功能预测, 对拟南芥GRAS家族基因在渗透和干旱胁迫过程中的应答模式进行了初步探索, 提出了一类响应渗透胁迫和干旱胁迫的拟南芥GRAS家族基因。以SCL13为例, 利用基因芯片相关性和GO分析, 对其在渗透胁迫信号转导过程中可能的调控机制进行了预测和分析。这一研究将为阐明GRAS家族基因参与水分胁迫的分子机制提供新的思路, 同时也为植物抗逆分子育种提供候选基因。     

海草形态、生长的种间差异及其相关生长关系

海草是海洋沉水高等植物,属典型的根茎克隆植物.根状茎直径和分株重分别是其个体大小的第一、二表征指标,个体大小是海草重要的种间识别特征.对海草6个形态构件指标和18个生长指标的综合分析表明:果实大小、单株叶面积、分株重具有显著的种特异性;分株发出的时间差、水平根状茎分枝率、叶年产量、分株寿命和垂直茎分枝率是海草种特异性最强的5个生长动态指标,海草生长动态的差异主要体现在克隆生长能力强弱和分株生活史长短上;大海草趋于游击型克隆构型,而小海草则趋于密集型,但小海草Cymodocea nodosa例外;大海草基株水平扩展能力较小海草差.海草个体大小与生长特征的相关生长关系表明:随个体的增大,海草在有机构件生长上表现出两相邻叶、相邻分株、相邻节发出的时间差延长,分株、叶、茎寿命延长的特点;在克隆生长水平上表现出根状茎节间长变短、延伸速率降低,分枝率和根状茎上年产分株数降低,分枝角度变小和间隔子增大的趋势;在克隆片段水平上表现为生理整合性增强;在种群层面则表现出生物量增大和种群密度降低的特点.因此,海草个体大小对其形态、生长特征、克隆构型、种群密度和生产力起到了决定作用.大小海草不同的形态、生长动态和克隆构型特征导致它们的生存策略及生态功能也不同,这一点可能对海草场修复基础理论研究具有一定的指示作用.     

Electrochemical biosensor for the detection of cauliflower mosaic virus 35 S gene sequences using lead sulfide nanoparticles as oligonucleotide labels

Lead sulfide (PbS) nanoparticles were synthesized in aqueous solution and used as oligonucleotide labels for electrochemical detection of the 35 S promoter from cauliflower mosaic virus (CaMV) sequence. The PbS nanoparticles were modified with mercaptoacetic acid and could easily be linked with CaMV 35 S oligonucleotide probe. Target DNA sequences were covalently linked on a mercaptoacetic acid self-assembled gold electrode, and DNA hybridization of target DNA with probe DNA was completed on the electrode surface. PbS nanoparticles anchored on the hybrids were dissolved in the solution by oxidation of HNO₃ and detected using a sensitive differential pulse anodic stripping voltammetric method. The detection results can be used to monitor the hybridization reaction. The CaMV 35 S target sequence was satisfactorily detected with the detection limit as 4.38 × 10⁻¹² mol/L (3σ). The established method extends nanoparticle-labeled electrochemical DNA analysis to specific sequences from genetically modified organisms with higher sensitivity and selectivity.     

禽病原性大肠杆菌iro和tsh基因缺失株的构建

通过SSH和SCOTS研究, 铁系统(Iro)和温度敏感性血凝素(Tsh)在禽病原性大肠杆菌(APEC)的感染中可能发挥重要作用。基因检测发现, 在243个禽源大肠杆菌分离株中, 有205株为iro+菌株, 其中高、中度和低致病株分别为89.8%(184/205)、8.8%(18/205)和1.5%(3/205); 有167株为tsh+菌株, 高、中度、低致病株分别为87.4%(146/167)、12.6%(21/167)和0%(0/167), 结果显示iro+或tsh+株大多数为高致病株。为了确定iro和tsh基因在APEC致病力中的作用, 以APEC E037株为基础, 通过自杀性载体分别构建了iro和tsh基因缺失突变株E037(Δiro)、E037(Δtsh)和E037(ΔiroΔtsh)。动物感染性试验表明, 突变株在鸡体内的繁殖能力和致病性均明显下降, 但两个基因的协同致病作用不显著。进一步证实Iro和Tsh为APEC重要的致病因子。