猪HK2基因的克隆及序列分析

为克隆猪己糖激酶2基因,分析其生物功能,采用已报道的人及小鼠HK2的cDNA序列为依据,利用电脑克隆策略获得的ESTs设计引物,扩增新的猪HK2基因cDNA序列。将PCR产物克隆测序,分析获得的核苷酸序列及其编码蛋白的特性,分离的开放阅读框全长2754bp,编码917个氨基酸,与人和小鼠的核苷酸序列同源率分别为90%和87%,与人和小鼠的氨基酸序列同源率分别为96%和94%,利用生物信息学软件分析出此蛋白的理化特性并预测了其蛋白结构,为进一步开展猪HK2基因的结构功能、表达调控的相关研究奠定了基础。     

猪HK2基因的克隆及序列分析

为克隆猪己糖激酶2基因,分析其生物功能,采用已报道的人及小鼠HK2的cDNA序列为依据,利用电脑克隆策略获得的ESTs设计引物,扩增新的猪HK2基因cDNA序列。将PCR产物克隆测序,分析获得的核苷酸序列及其编码蛋白的特性,分离的开放阅读框全长2754bp,编码917个氨基酸,与人和小鼠的核苷酸序列同源率分别为90%和87%,与人和小鼠的氨基酸序列同源率分别为96%和94%,利用生物信息学软件分析出此蛋白的理化特性并预测了其蛋白结构,为进一步开展猪HK2基因的结构功能、表达调控的相关研究奠定了基础。     

Porosity Distribution,Surface Area,and Morphology of Synthetic Potassium Zeolites: A SEM and N2 Adsorption Study

The morphology of a synthetic zeolite (1717-nm particle-size grains; pore width 10–15 nm) as well as its surface area and mesoporosity is described. Morphology investigated by field emission scanning electron microscopy has shown cuboid crystals. The full adsorption–desorption isotherms for N₂ gas measured volumetrically at 77 °K have shown type IV isotherm with type H₃ hysteresis loops. To study the reliable pore-size distribution, both adsorption and desorption, curves were used by fitting the data to several well-known adsorption models: Brunauer–Emmett–Teller (BET), Barret, Joyner, and Halenda (BJH), Dollimore and Heal (D-H), Horvath–Kawazoe, and the density functional theory (DFT).     

Influence of heating and drying history on micropores in dry wood

To investigate the influence of heating and drying history on the microstructure of dry wood, in addition to the dynamic viscoelastic properties, CO₂ adsorption onto dry wood at ice.water temperature (273 K) was measured, and the micropore size distribution was obtained using the Horvath-Kawazoe (HK) method. Micropores smaller than 0.6 nm exist in the microstructures of dry wood, and they decreased with elevating out-gassing temperature and increased again after rewetting and drying. Dry wood subjected to higher temperatures showed larger dynamic elastic modulus (E′) and smaller loss modulus (E″). This is interpreted as the result of the modification at higher temperature of the instability caused by drying. Drying history influenced the number of micropores smaller than 0.6 nm in dry wood not subjected to high temperature, although the difference in the number of micropores resulting from the drying history decreased with increasing out-gassing temperature. A larger number of micropores smaller than 0.6 nm exist in the microstructure of dry wood in more unstable states, corresponding to smaller E′ and larger E″ than in the stable state. Consequently, unstable states are considered to result from the existence of temporary micropores in the microstructures of dry wood, probably in lignin. Part of this report was presented at the 55th Annual Meeting of the Japan Wood Research Society, Kyoto, March 2005, and at the 56th Annual Meeting of the Japan Wood Research Society, Akita, August 2006     

Changes in micropores in dry wood with elapsed time in the environment

To investigate the changes in microstructures of wood with elapsed time in the environment, CO₂ adsorption onto dry wood was measured at ice-water temperature (273 K) for samples aged from 0.1 years to over 1000 years. The micropore size distribution was obtained using the Horvath-Kawazoe method. Micropores smaller than 0.6 nm in wood decreased in number with elapsed time in the environment, and a negative correlation was found between cumulative pore volume for pores smaller than 0.6 nm and elapsed time in the environment. Cumulative pore volume in the 1000-year sample was almost half of that in the 0.1- year sample. Micropores smaller than 0.6 nm in wood with a few decades or more of elapsed time increased in number after rewetting and drying. Consequently, microstructures of wood with longer time elapsed in the environment were considered to be more stable, because of longer-term thermal motion and possibly more repeated moisture adsorption and desorption and/or temperature variation in the environment.     

温度对蜕壳后期三疣梭子蟹能量代谢酶活力的影响

为研究温度对三疣梭子蟹雌蟹钙化过程中丙酮酸激酶(PK)、己糖激酶(HK)和琥珀酸脱氢酶(SDH)活力的影响,在实验室条件下设18、22、26 ℃ 3 个温度水平,分别在梭子蟹蜕壳后1、2、8、16 h及完全硬化5个时间点取样,测定其肝胰脏、肌肉和鳃中HK、PK、SDH的活力。结果显示:①与22 ℃条件下相比,高温(26 ℃)和低温(18 ℃)下三疣梭子蟹肌肉中PK的活力在蜕壳后1～8 h明显升高,说明高温和低温条件下梭子蟹肌肉中的糖酵解速率加大;而肝胰脏和鳃中的PK活力的峰值出现在梭子蟹蜕壳后1～2 h,表明蜕壳后1～2 h这2种组织中有较大的糖酵解速率;②3个温度水平下,梭子蟹肝胰脏中HK的活力随钙化过程的进行而降低,蜕壳后1～16 h的活力显著高于完全钙化时的活力,说明蜕壳后的梭子蟹肝胰脏对葡萄糖的利用能力逐渐减弱,而其鳃中HK活力的变化趋势与肝胰脏中相反,说明梭子蟹的鳃对葡萄糖的利用能力随钙化过程的进行而提高,这种差异可能与不同组织在梭子蟹钙化过程中对葡萄糖的利用能力不同有关;③梭子蟹肝胰脏中SDH的活力在3个实验温度下蜕壳后16 h内呈逐渐升高的趋势,说明肝胰脏在钙化过程中三羧酸循环作用逐渐加强。此外,其肌肉和鳃组织中SDH的活力在3个实验温度下的峰值均出现在前8小时,并且低温(18 ℃)的峰值明显低于高温(26 ℃),说明在这段钙化时间中,相对于低温而言,梭子蟹的肌肉和鳃中有氧代谢的加强在高温条件下更显著。在钙化过程中,低温条件下三疣梭子蟹的糖酵解作用没有明显的变化,而三羧酸循环有较明显的减弱趋势,其中三疣梭子蟹肌肉和鳃中SDH的活力明显低于高温条件,表明低温确实可以延长其蜕壳后的软壳时间。     

Immunoregulation of fish nonspecific cytotoxic cell activity by retinolacetate but not poly I:CImmunomodulation de l'activité cytotoxique non spécifique de cellules de poisson par le rétinolacétate mais non par le poly I:C

Nonspecific cytotoxic cells (NCC) from fish (Ictalurus punctatus) were treated with different concentrations of retinolacetate and poly I:C. Both in vitro and in vivo experiments were conducted. Retinolacetate significantly increased NCC activity against chromium-51 labeled human B-cell lymphoma target cells (NC-37). Preincubation (treatment prior to adding the labeled target cells) of NCC for 4 to 8 h in 10^?3 to 10^?12 M concentration of retinolacetate produced significant increases in NCC activity compared to treatment during the killing assay only. Similar experiments with different concentrations of poly I:C had no NCC augmenting effects when tested by adding poly I:C either during preincubation periods or during the cytotoxicity assay. Retinolacetate probably produces positive modulation of cytotoxicity by increasing the killing effectiveness of individual NCC, rather than recruiting larger numbers of cytolytic cells. In vivo studies were also conducted by injecting catfish (i.p.) with 1 ×, 3 × and 5 × the daily recommended vitamin A dosages and determining NCC activity after 24, 48 and 72 h treatment. The 1 × dose significantly increased NCC activity at 72 h. This increase was not transient because NCC activity after 33–37 days' treatment was significantly higher than controls in the 1 ×, 2 × and 3 × groups. Intraperitoneal injections of fish with poly I:C produced no significant increases in NCC activity at 24 or 72 h post-inoculation.     

Micropore structure of wood: Change in micropore structure accompanied by delignification

In our preceding study, we clarified that liquids having similar molecular sizes to ethanol were mainly adsorbed onto lignin among the major constituents of wood. This suggests that most micropores or adsorption sites loosely hydrogen-bonded to each other, which are accessible to these liquids, exist in lignin. In the present study, to examine micropores in wood and lignin, micropore distribution was measured by CO₂ gas adsorption at ice-water temperature (273 K). Dry samples prepared by gradual delignification from wood meal were used as adsorbents. The pore-size distributions were determined by analyzing adsorption isotherms using the Horvath-Kawazoe method. It was found that the number of micropores decreased with the decrease in residual lignin, and micropores were hardly found in cellulose and hemicellulose. It is considered that most micropores smaller than 0.6 nm in wood exist in lignin.     

蒺藜状苜蓿HK1的基因组解析

利用生物信息学的方法,从蒺藜状苜蓿的基因组中解析得到一个与拟南芥渗透胁迫感受器基因AtHK1同源的基因,命名为MtHK1,并对它的基因结构、蛋白基序、启动子区的顺式元件和基因的遗传进化进行了分析。结果显示,蒺藜状苜蓿基因MtHK1编码的蛋白具有组氨酸蛋白激酶的结构特征,而且在进化上与拟南芥的AtHK1分属同一个类群。从基因顺式反应元件分析来看,蒺藜状苜蓿基因MtHK1具有数个响应不同逆境和激素的顺式元件,预示这个基因在逆境响应和植物生长发育过程中具有一定功能。本文为下一步深入研究这个基因的生物学功能奠定了基础。     

一个番茄细胞分裂素受体类激酶基因的生物信息学分析

利用生物信息学的方法从番茄的基因组中解析得到一个与拟南芥渗透胁迫相关的细胞分裂素受体类激酶HK1高度同源的基因,命名为LeHK1,并对它的序列结构、蛋白基序、启动子区的顺式元件和基因的遗传进化进行了分析。结果显示,番茄基因LeHK1编码的蛋白具有组氨酸蛋白激酶的结构特征,而且在进化上与拟南芥的渗透胁迫感受基因AtHK1分属同一个类群。从基因顺式反应元件分析来看,番茄基因LeHK1具有多个响应不同环境信号如热、干旱、激素信号的顺式元件,预示这个基因在环境响应和植物生长发育过程中具有重要功能。