大肠杆菌表达重组人生长激素的纯化

目的 :获得具有生物学活性的重组人生长激素 (rhGH)。方法 :PBV -GH/DH5α菌体经超声破菌、反复洗涤后获得包涵体。将包涵体变性、复性 ,用硫酸铵盐析 ,离子交换层析和凝胶层析进行纯化。产物经SDS -PAGE、HPLC、N末端 15个氨基酸序列检测验证。结果 :终产物rhGH纯度达 98.2 % ,比活性大于 3.0IU/mg。分子量为 2 2kDa ,N末端氨基酸序列与DNA序列推导的氨基酸序列完全一致。结论 :从自构建的PBV -GH/DH5α工程菌中获得高纯度、高活性重组人生长激素。其纯化工艺为中试生产提供可靠依据。     

低镁胁迫对低温下黄瓜幼苗光合特性和抗氧化系统的影响

以‘津优3号’黄瓜幼苗为试材,以Hoagland全营养液处理为对照(CK),研究低镁(30％Mg)胁迫对低温(昼/夜温度12℃/8℃)下黄瓜幼苗光合特性和抗氧化系统的影响.结果表明:低温下30％Mg处理黄瓜幼苗叶片Mg含量显著低于CK,而根中Mg含量与CK差异不显著.随着低温胁迫时间的延长,黄瓜幼苗叶片的叶绿素含量、净光合速率(Pn)、气孔导度(gs)和羧化效率(CE)逐渐降低,胞间CO2浓度(Ci)趋于升高.与CK相比,低温下30％ Mg处理叶片叶绿素含量、Pn、gs和CE显著降低,C-变化不大,叶绿体膜损伤严重,叶绿体数、基粒数和片层数较少,淀粉粒数增加,淀粉粒较长,丙二醛(MDA)含量升高,而超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)和谷胱甘肽还原酶(GR)活性降低.可见,低温下镁运输受阻引起的缺镁是叶片失绿的主要原因;低温引起Pn降低的主要原因是非气孔限制,低镁胁迫会加大低温对黄瓜Pn的影响,而由此引起的Pn降低的主要原因是气孔限制.     

浙江省国家重点风景名胜区楠溪江两栖动物资源调查与区系分析

为了解浙江省永嘉县国家重点风景名胜区楠溪江野生动物资源现状,促进风景区的有效管理和建设,于2008年5月、8月、9月、10月和2009年5月对楠溪江流域的两栖动物进行野外调查.调查结果表明,楠溪江流域共计有两栖类24种,隶属于2目7科,中国特有种13种,其中福建大头蛙和天台粗皮蛙是温州地区的新纪录种.区系组成以东洋界华中华南区共有种为主,东洋界占明显优势,达22种,占物种总数的91.7%.     

鹿蹄草植物的研究与应用

对鹿蹄草植物的自然资源分布、化学成分、药理作用、及其应用的相关研究做了综述。该植物在世界各地广泛分布、含有鹿蹄草素等多种化学成分,药理研究证明其具有抗菌、抗病毒、消炎、镇痛作用、对心脑血管系统等作用,主要应用在医药工业、食品工业和园林绿化面。最后对该属植物的研究前景做了展望。     

G protein beta gamma subunit interaction with the dynein light-chain component Tctex-1 regulates neurite outgrowth

Tctex-1, a light-chain component of the cytoplasmic dynein motor complex, can function independently of dynein to regulate multiple steps in neuronal development. However, how dynein-associated and dynein-free pools of Tctex-1 are maintained in the cell is not known. Tctex-1 was recently identified as a Gbetagamma-binding protein and shown to be identical to the receptor-independent activator of G protein signaling AGS2. We propose a novel role for the interaction of Gbetagamma with Tctex-1 in neurite outgrowth. Ectopic expression of either Tctex-1 or Gbetagamma promotes neurite outgrowth whereas interfering with their function inhibits neuritogenesis. Using embryonic mouse brain extracts, we demonstrate an endogenous Gbetagamma-Tctex-1 complex and show that Gbetagamma co-segregates with dynein-free fractions of Tctex-1. Furthermore, Gbeta competes with the dynein intermediate chain for binding to Tctex-1, regulating assembly of Tctex-1 into the dynein motor complex. We propose that Tctex-1 is a novel effector of Gbetagamma, and that Gbetagamma-Tctex-1 complex plays a key role in the dynein-independent function of Tctex-1 in regulating neurite outgrowth in primary hippocampal neurons, most likely by modulating actin and microtubule dynamics.     

南方鲇的营养学研究:饲料中大豆蛋白水平对消化率及摄食率的影响

以23.78±0.09g的南方鲇为实验对象,以Cr2O3作为测定饲料消化率的指示物,以大豆蛋白分别替代0%、13%、26%、39%、52%和65%的鱼粉蛋白,配制成6种等氮(粗蛋白48%)、等能(总能20KJ/g)的饲料,用于探讨饲料中大豆蛋白替代鱼粉蛋白的比例对南方鲇消化率及摄食率的影响。实验在27.5±0.02℃水温条件下进行6周,溶氧维持在5mg/L以上,光制为14L∶10D。结果表明,饲料中不同比例的鱼粉蛋白与大豆蛋白对南方鲇的消化率及摄食率均存在显著影响(P<0.05)。其中南方鲇对于物质、蛋白质、脂肪和能量的消化率随着大豆蛋白替代率的提高而逐步下降,当替代量为65%时,其消化率显著低于其余各组(P<0.05)。而南方鲇的摄食率均随着饲料中大豆粉添加量的升高而升高,当大豆蛋白替代鱼粉蛋白超过39%时,摄食率显著高于其余各组(P<0.05)。通过分析认为,消化率是限制南方鲇利用大豆粉的重要因素,而摄食率不是影响其利用率的主要原因。     

分子伴侣GroE系统能量传递机制的研究

用SwissPDBViewer软件对分子伴侣GroE系统与底物的相互作用进行了模拟 ,结果表明 :GroEL顶端结构域在GroES和靶蛋白结合之后发生了明显的变化 ;GroEL的cis环上有与三磷酸腺苷ATP相结合的位点 ,ATP水解之后形成的ADP与活性中心的残基相结合 ,而这种结合除导致残基Thr30的构型发生了变化之外 ,其它残基的空间位置和构型基本保持不变 ,暗示其它残基在能量传递过程中形成了刚性骨架 ,而与ADP分子磷酸键结合的残基Thr30则是能量传递的力点。     

绵羊GDF9和BMP15基因多态性检测

以绵羊GDF9基因FecG^H突变和BMP15基因FecX^B和FecX^G突变为候选基因,采用PCR—RFLP方法研究其在湖羊、夏洛来、陶赛特、萨福克、中国美利奴肉用多胎品系、中国美利奴羊和罗米丽羊7个品种中的多态性．结果发现,湖羊群体中存在GDF和BMP15基因的FecG^H和FecX^B突变,但发生率极低,分别为0．645％(2／310)和0．968％(3／310)：而其它品种中则没有发现GDF9和BMP15基因的相应突变．这一发现对于建立绵羊基因标记辅助选择方法具有重要意义．     

Perturbing the ubiquitin pathway reveals how mitosis is hijacked to denucleate and regulate cell proliferation and differentiation in vivo

Background

The eye lens presents a unique opportunity to explore roles for specific molecules in cell proliferation, differentiation and development because cells remain in place throughout life and, like red blood cells and keratinocytes, they go through the most extreme differentiation, including removal of nuclei and cessation of protein synthesis. Ubiquitination controls many critical cellular processes, most of which require specific lysines on ubiquitin (Ub). Of the 7 lysines (K) least is known about effects of modification of K6.

Methodology and Principal Findings

We replaced K6 with tryptophan (W) because K6 is the most readily modified K and W is the most structurally similar residue to biotin. The backbone of K6W-Ub is indistinguishable from that of Wt-Ub. K6W-Ub is effectively conjugated and deconjugated but the conjugates are not degraded via the ubiquitin proteasome pathways (UPP). Expression of K6W-ubiquitin in the lens and lens cells results in accumulation of intracellular aggregates and also slows cell proliferation and the differentiation program, including expression of lens specific proteins, differentiation of epithelial cells into fibers, achieving proper fiber cell morphology, and removal of nuclei. The latter is critical for transparency, but the mechanism by which cell nuclei are removed has remained an age old enigma. This was also solved by expressing K6W-Ub. p27^kip, a UPP substrate accumulates in lenses which express K6W-Ub. This precludes phosphorylation of nuclear lamin by the mitotic kinase, a prerequisite for disassembly of the nuclear membrane. Thus the nucleus remains intact and DNAseIIβ neither gains entry to the nucleus nor degrades the DNA. These results could not be obtained using chemical proteasome inhibitors that cannot be directed to specific tissues.

Conclusions and Significance

K6W-Ub provides a novel, genetic means to study functions of the UPP because it can be targeted to specific cells and tissues. A fully functional UPP is required to execute most stages of lens differentiation, specifically removal of cell nuclei. In the absence of a functional UPP, small aggregate prone, cataractous lenses are formed.     

卡托普利对高血压大鼠动脉血管平滑肌细胞内质网相关因子葡萄糖调节蛋白78和C/EBP同源蛋白表达的影响

目的:研究卡托普利对早期高血压大鼠动脉血管平滑肌细胞内质网相关因子葡萄糖调节蛋白78(glucose-regulated protein of 78kd,GRPT8)和C/EBP同源蛋白(CAAT/enhancer binding protein homologous protein,CHOP)表达的影响.方法:将18只成年雄性Sprague-Dawley(SD)大鼠随机分为对照组、模型组和卡托普利组(n=6),模型组和卡托普利组均采用大鼠腹主动脉结扎建立高血压大鼠模型.4周后测量血压,应用免疫组化方法检测主动脉血管平滑肌细胞GRP78和CHOP的表达;缺口末端标记法(TUNEL)检测细胞凋亡.结果:(1)模型组平均动脉压(mean arterial blood pressure,MAP)明显增加,卡托普利组血压低于模型组,高于对照组,差异具有统计学意义(P<0.05);(2)模型组内质网因子GRP78、CHOP表达均增加,卡托普利组GRP78、CHOP表达低于模型组,高于对照组,差异具有统计学意义(P<0.01);(3)模型组血管平滑肌细胞凋亡率减少,卡托普利组血管平滑肌细胞凋亡率高于模型组,低于对照组,差异具有统计学意义(P<0.01).结论:卡托普利可降低高血压大鼠动脉血管平滑肌细胞GRP78和CHOP的表达,增加细胞凋亡.可能与其减弱高血压所致内质网反应,维持血管平滑肌细胞的增殖/凋亡平衡有关.