植物高赖氨酸蛋白质的分离和纯化及其N端序列核苷酸探针的合成

禾谷类作物水稻和小麦是人们的主要植物性食物来源．而这些作物种子蛋白质中人类不能合成的必需氨基酸含量不平衡，造成了优质蛋白质的缺乏和人体对蛋白质利用的极大浪费．大约有二分之一的谷类种子蛋白质和四分之一的大豆蛋白质不能被合理地利用。大多数禾本科作物包括水稻和小麦的种子蛋白质中第一限制性氨基酸是赖氨酸，纠正其不平衡现象可大大提商蛋白质的营养价值。本研究在高赖氨酸植物种的筛选、高赖氨酸种子储藏蛋白质的纯化及其基因的分离等方面开展了工作。 选用与禾本科亲缘关系较远的8个植物种为研究材料，它们分别属于榛科，十字花科、胡桃科、豆科、胡麻科和松科。氨基酸组分分析确定豆科和十字花科的三个植物种赖氨酸含量在5.5%以上，其中豆科植物四棱豆(Pso phocarpus tetragonolobus)种子全蛋白赖氨酸含量达7.9%．用5种提取液提取了四棱豆种子的清蛋白、球蛋白和全蛋白。经测定发现0.025M Tris.HCl(pH7.4)提取液提取的清蛋白赖氨酸含量高最．通过自然胶电泳，SDS-PAGB电泳，非变牲IEF和变性IEF／SDS双向电泳，对四棱豆种子清蛋白进行了定性研究。用变性IEF/ SDS双向电泳分析出60多种蛋白质和蛋白质亚基及多肽。研究中改进了等电聚焦电泳纯化蛋白质的方法，经处理的胶板显现出清晰的蛋白质带型，不需染色即可确定带的位置，从切下的胶条中洗脱的蛋白质，其纯度达到双向电泳纯和HPLC纯。用三种电溶方法(SDS-PAGE非变性IEF，变性IEF)纯化出三十一种蛋白质或多肽分子。分别进行了分子量确定和氨基酸组分分析，发现了一个赖氨酸含量高达11.4%的蛋白质，其分子量为18KD，并制备了该蛋白质的抗体，测定了18KD蛋白质N端30个氨基酸残基的顺序，根据这一顺序设计合成了一组17个核苷酸的基因探针．经鉴定单链DNA探针的纯度和总量达到了设计要求。用尿素法与CTAB法结合提取了四棱豆幼苗核基因组DNA，其分子量在50Kb以上，达到了构建GenormicDNA文库的要求．用bamHI EcoRI和HindⅢ三种酶切割提取的DNA，得到了分子量大小不同的片段。 对四棱豆种子蛋白质的定性、高赖氨酸蛋白质的纯化、18KD蛋白N端序列分析及寡核苷酸探针的合成以及GcnomicDNA的提取与酶切，尚未见有资料报道．这些工作为克隆高赖氨酸基因打下了良好的基础，对改良禾本科作物蛋白质品质意义深远．

细胞色素b基因序列与11种熊蜂的系统进化

通过比较一段357bp的细胞色素b的序列,分析了熊蜂属5亚属11种熊蜂系统发育关系。5亚属包括Bombus(5种)、Thoracobombus(3种)、Mendacibombus(1种)、Fervidobombus(1种)和Pyrobombus(1种)。该序列有65个单变异多态位点和71个简约信息多态位点,翻译成119个氨基酸序列后有45个氨基酸变异位点。根据P-距离构建的邻接树(NJ tree)和最大简约树(MP tree)都显示同样的结果:Mendacibombus(B. avinovielllus)分化最早;Fervidobombus(B. pensylvanicus)次之;Pyrobombus(B. impatiens)和Bombus形成姊妹群;Bombus亚属是单系群,其中B. ignitus在所研究的5个种中分化最早。

Detour behavior changes associated with prenatal morphine exposure in 11-day-old chicks

The central nervous system exhibits remarkable plasticity in early life. Prenatal morphine exposure may induce adverse behavioral effects on the neonate and the developing offspring. In the present study, we investigated the effect of prenatal morphine exposure (daily from embryonic days 12-16, 20 mg/kg) on 11-day-old chicks using two forms of spatial paradigms: one trial detour behavior task in which animals must bypass an obstacle to reach the desired goal without any training and detour learning task which required several trials of training to reach the detour criterion. The results showed that, on the condition that chicks could successfully detour in the first trial, morphine exposed chicks exhibited longer detour latency to finish the task, coupled by a preference for turning right versus turning left. In contrast, no significant difference in learning and memory was found in detour learning task between morphine exposed chicks and saline chicks. These findings suggest specific behavioral changes associated with prenatal exposure to opioids during mid to late gestation, also raise attention to the possible health hazard from pregnancy drug use in everyday life. (C) 2010 ISDN. Published by Elsevier Ltd. All rights reserved.

HSF2 mRNA 在热应激和大剂量11酸睾酮诱导恒河猴生精细胞凋亡中的表达变化

为了探讨 HSF2 mRNA 在热应激和超生理剂量睾酮诱导恒河猴生精细胞凋亡中的表达变化, 作者建立了手术诱导单侧隐睾和注射大剂量11酸睾酮(TU)恒河猴动物模型, 应用3′末端标记分析(TUNEL)和原位杂交方法, 检测睾丸细胞的凋亡信号和 HSF2的表达变化. TUNEL 结果显示热应激和超生理剂量睾酮能够诱导生精细胞出现凋亡信号, 它分别于处理后第5天和第30天达到最强, 表明热应激和睾酮干扰精子发生可能是通过生精细胞凋亡的方式来实现的. HSF2 mRNA 水平在生精细胞凋亡早期(凋亡信号达到最强以前)略有降低, 而在凋亡高峰期之后其表达急剧下降. Hsf2基因与作者以前研究的 Hsp70-2基因的表达具有时间上的相关性, 表明 HSF2蛋白可能调控 Hsp70-2基因的表达, 而且 HSF2可能通过多种方式影响精子的发生以及抑制生精细胞的凋亡。

P16蛋白和生精细胞凋亡对热压和11酸睾酮诱导的恒河猴无精子症和少精子症中的作用

探讨了P16蛋白和生精细胞凋亡在热压和11酸睾酮诱导恒河猴无精子症和少精子症中作用间的关系。3^末端标记分析(TUNEL)结果显示热应激和超生理剂量睾酮能够诱导生精细胞出现凋亡信号,它分别于处理后d5和d30达到最强。免疫组化结果显示,热压或TU主要诱导精原细胞和其它生精细胞以及Sertoli细胞P16的表达。P16蛋白的表达在生精细胞凋亡晚期,即隐睾手术d10或注射TUd60后迅速升高并维持高表达,该蛋白在生精细胞凋亡晚期可能通过抑制精原细胞的有丝分裂,扰乱正常的精子发生。

异育银鲫对鱼粉等11种饲料中磷的利用率

于1988年,以三氧化二铬间接指示法,在水温22±2℃条件下,测定了平均体重为55±5g/尾的异育银鲫对鱼粉、肉骨粉、芝麻饼、虾粉、骨粉、菜籽饼、玉米粉、脱氟磷酸氢钙、磷酸二氢钙、磷酸氢钙、磷酸钙的表观消化吸收率。结果表明:异育银鲫对第一、第二、第三磷酸钙的消化吸收率是不同的,分别为81％,65％和32％,此顺序与其溶解性由大到小的顺序一致;在所测试的八种商品饲料中,对骨粉、虾粉和菜籽饼中磷的表观消化吸收率较高,分别为65％,68％和65％;对鱼粉、芝麻饼、肉骨粉三种原料中磷的表观消化吸收率较低,分别是1

Effects of polarization field on formation of two-dimensional electron gas in (0001) and (11(2)over-bar0) plane AlGaN/GaN heterostructures

Photoluminescence (PL) and temperature-dependent Hall effect measurements were carried out in (0001) and (11 (2) over bar0) AlGaN/GaN heterostructures grown on sapphire substrates by metalorganic chemical vapor deposition. There are strong spontaneous and piezoelectric electric fields (SPF) along the growth orientation of the (0001) AlGaN/GaN heterostructures. At the same time there are no corresponding SPF along that of the (1120) AlGaN/GaN. A strong PL peak related to the recombination between two-dimensional electron gas (2DEG) and photoexcited holes was observed at 3.258 eV at room temperature in (0001) AlGaN/GaN heterointerfaces while no corresponding PL peak was observed in (11 (2) over bar0). The existence of a 2DEG was observed in (0001) AlGaN/GaN multi-layers with a mobility saturated at 6000 cm(2)/V s below 80 K, whereas a much lower mobility was measured in (11 (2) over bar0). These results indicated that the SPF was the main element to cause the high mobility and high sheet-electron-density 2DEG in AlGaN/GaN heterostructures. (C) 2004 Elsevier B.V. All rights reserved.

Luminescence study of (11(2)over-bar0) GaN film grown by metalorganic chemical-vapor deposition

Investigations on photoluminescence properties of (11 (2) over bar0) GaN grown on (1 (1) over bar 02) Al2O3 substrate by metalorganic chemical-vapor deposition are reported. Several emission lines not reported before are observed at low temperature. The sharp peak at 3.359 eV is attributed to the exciton bound to the neutral acceptor. Another peak at 3.310 eV represents a free-to-bound, probably a free electron-to-acceptor, transition. The 3.241 and 3.170 eV lines are interpreted as phonon replica lines of the 3.310 eV line. The phonon energy is 70 meV, consistent with the energy of transverse optical E-1 phonon. The optical properties of the lines are analyzed. (C) 2003 American Institute of Physics.