The 65-kDa cytosolic protein associated with warm temperature acclimation in goldfish,Carassius auratus

Goldfish Carassius auratus were acclimated to either 10 or 30°C for a minimum of 5 weeks. A 65-kDa protein specific to warm-temperature-acclimated fish was extracted from the gel with 70% formic acid after two-dimensional electrophoresis of the muscle cytoplasmic protein fraction. The 65-kDa protein thus prepared to homogeneity was used to raise specific antibodies in rabbit by conventional methods. The antibody produced exhibited specific reaction with a protein having the same molecular weight from brain and liver tissue, suggesting that the 65-kDa protein is a ubiquitous cytosolic component in warm-acclimated goldfish. When water temperature was increased from 20 to 30°C over a 20-h period, a prominent amount of the 65-kDa protein was observed in muscle tissue extracts within 5 days of additional rearing; this was demonstrated by immunoblotting with the specific antibody. The N-terminal amino acid sequence of the 65-kDa protein was determined as Asp-Glu-Pro-Gln-Gly-His-Gln-His (or Asp)-Glu-Leu, differing from that of a family of known heat-shock proteins having about 70 kDa in molecular mass (hsp 70). No interaction between ATP and the 65-kDa protein revealed by ATP-agarose affinity chromatography further confirmed the different properties of the 65-kDa protein from those of hsp 70.Abbreviations ATP adenosine 5-triphosphate - hsp heat-shock protein(s) - IgG immunoglobulin G - mRNA messenger ribonucleic acid - PMSF phenylmethylsulphonyl fluoride - PVDF polyvinylidene difluoride - SDS sodium dodecyl sulphate - SDS-PAGE SDS-polyacrylamide gel electrophoresis     

人白细胞介素－3在大肠杆菌中的高效表达

人白细胞介素－３（ｈｕｍａｎＩｎｔｅｒｌｅｕｋｉｎ３,ｈＩＬ－３）是一种造血前体细胞早期分化的关键调节因子。用ＰＣＲ方法从人Ｔ淋巴细胞ｃＤＮＡ文库中扩增出０．４４ｋｂ的ＤＮＡ片段,并克隆入ｐＵＣ１９载体中。经ＤＮＡ序列测定,确定０．４４ｋｂ的ＰＣＲ产物合完整的编码人白细胞介素－３成熟蛋白ｃＤＮＡ序列,并在信号肽与成熟蛋白编码序列之间通过突变引入了限制性内切酶位点和ＡＴＧ起始密码。构建的ＰＬ启动子控制下的ｈＩＬ－３ｃＤＮＡ表达质粒,转入大肠杆菌Ｔａｐ１０６,经４２℃热诱导,获得ｈＩＬ－３的表达产物。ＳＤＳ－ＰＡＧＥ电泳显示表达产物为１５ｋｄ,约占细菌总蛋白的１５％。表达产物经ＥＬＩＳＡ和Ｗｅｓｔｅｒｎ－ｂｌｏｔ验证。ｈＩＬ－３表达产物在细胞内形成包涵体,纯化包涵体,使产物纯度提高到７０．８％,产物复性后,能明显促进ｈＩＬ－３依赖细胞株生长,具有明显的生物活性。产物转移到ＰＶＤＦ膜后进行Ｎ端序列分析,Ｎ端１６个氨基酸正确。     

Functions of milk protein gene 5′ flanking regions on human growth hormone gene

Fragments containing 5′ flanking regions of four bovine milk protein genes—alpha lactalbumin (bαLA), alpha S₁ casein (bαS₁CN), beta casein (bβCN), kappa casein (b_kCN)—and mouse whey acidic protein (mWAP) gene were prepared by PCR and ligated to human growth hormone (hGH) gene. These recombinant DNAs were microinjected into rat embryos to produce transgenic rats, and the functions of the 5′ regions to direct secretion of hGH in the milk were tested. Although milk was obtained only in 5 of 19 mWAP/hGH rat lines, more than two-thirds of the rats carrying the other four DNAs produced milk. More than 80% of the lactated rats carrying bαLA/, bβCN/, and mWAP/hGH, and 33% of the laclated bαS₁CN/hGH rats secreted detectable amounts of hGH (> 0.05 μg/ml) in the milk. In some rats, the hGH concentrations in the milk were comparable to or more than that of the corresponding milk protein in bovine milk. The ranges of hGH concentrations in the milk of bαLA/, bβCN/, bαS₁CN/, and mWAP/hGH rats were 1.13–4,360 μg/ml, 0.11–10,900 μg/ml, 86.8–6,480 μg/ml, and 6.87–151 μg/ml, respectively. HGH was also detected in the sera of these rats, and some abnormalities of growth and reproduction were observed. All but one virgin mWAP/hGH rat secreted up to 0.0722 μg/ml of hGH in the serum, and more than half of them showed abnormal fat accumulations at their abdomen. None of the bαCN/hGH rats secreted detectable amount of hGH into their milk, whereas 8 of the 11 lines secreted hGH into their sera. For the production of hGH in transgenic rat milk, the 5′ region of bαS₁CN was shown most suitable, because the bαS₁CN/hGH rat secreted > 6,000 μg/ml of hGH into the milk and could be reproduced. © 1994 Wiley-Liss, Inc.     

Morphological and biological characteristics of the Korean population of Leptoglossus occidentalis Heidemann (Heteroptera: Coreidae), an invasive insect pest of conifer cones

In this study, we investigated the morphological and biological characteristics of a serious insect pest of conifer cones, the western conifer seed bug (WCSB), Leptoglossus occidentalis Heidemann (Heteroptera: Coreidae), in Korea. WCSB adults shared external morphology with overseas populations, including zig-zag fascia on the coria, dilation of the hind tibiae, and color pattern on the abdominal dorsum. The average body length and width of WCSB adults were 17.20 and 4.85 mm, respectively, in males and 19.05 and 5.55 mm in females. The average body weights were 111.27 and 169.89 mg in adult males and females, respectively. On average, eggs were 2.12 mm long and 1.17 mm wide. The average body lengths of the first- to fifth-instar nymphs were 3.07, 4.76, 8.77, 13.56, and 15.53 mm, respectively. For biological characteristics, developmental durations of the egg and five nymphal instar stages were 8.6, 3.7, 7.6, 5.8, 5.8, and 9.1 days, respectively, in laboratory conditions. Nymphal mortality was 47.5% in total. The average life expectancy of WCSB adults was 58.2 and 63.9 days in males and females, respectively. Oviposition of the adult females started 23.4 days after emergence, and the total number of eggs laid by a single adult female was 68.6 on average. The results of this study offer basic morphological and biological information that can be used to design an effective control strategy to manage the WCSB population in Korea.     

泉州常绿阔叶林的特征与分布

泉州的常绿阔叶林包括南亚热带雨林和南亚热带山地照叶林。通过对这两种植被类型的区分成分、生活型、叶的性质以及群落的结构与分布的分析,说明海拔450m以下具有较明显的以厚壳桂、红楠等为优势种的南亚热带雨林的特征和景观;海拔450～900m主要以米槠等拷类为优势种的群落,但仍有南亚热带雨林的某些特征;海拔900～1400m以甜槠为主要建群种的群落,属于中亚热带照叶林的群落特征。     

一株C3形态多重耐药大肠杆菌噬菌体生物学特性和基因组分析

【目的】多重耐药菌的出现对公共卫生安全构成严重威胁,本研究分离多重耐药大肠杆菌噬菌体,研究其生物学特性和基因组特征,为耐药菌的噬菌体疗法提供理论依据。【方法】使用双层平板法从污水样本中分离纯化大肠杆菌噬菌体;磷钨酸染色后通过透射电镜观察形态;测定其宿主范围,测定温度和pH稳定性、一步生长曲线和体外抑菌效果等生物学特性;体内抑菌试验评估噬菌体对多重耐药大肠杆菌N1203-1Af感染的大蜡螟幼虫的保护作用;基于全基因组测序对其基因组特点进行分析。【结果】本研究分离共得到5株大肠杆菌噬菌体,分别命名为pEC-S163-2.1、pEC-S163-2.2、pEC-M1167-5Ar.1、pEC-m1291-2Dr.1和pEC-N1203-2Af.1;电镜结果显示噬菌体pEC-N1203-2Af.1属于短尾噬菌体中罕见的C3形态型,头部较长,长是宽的2–3倍;pEC-N1203-2Af.1可裂解受试15株大肠杆菌中的3株;感染10 min后进入指数增长期,–20-50℃、pH值为4.0–10.0的环境下均能够保持稳定活性;大蜡螟幼虫感染大肠杆菌N1203-2Af后噬菌体pEC-N1203-2Af....     

尕海湿地草甸土退化过程土壤氮矿化演变特征

氮矿化是生态系统循环的重要环节之一,影响着生态系统功能和氮素生物地球化学循环,因此研究高寒湿地退化过程中土壤氮矿化演变特征,对揭示气候变化和人为活动干扰背景下的湿地土壤氮素循环过程具有重要意义。以尕海湿地4种不同退化梯度(未退化、轻度退化、中度退化、重度退化)土壤为研究对象,采用野外树脂芯原位培养方法,通过对植物生长季不同生长阶段(生长初期、生长盛期、枯萎期)土壤氮素矿化作用研究,分析湿地退化演替过程中土壤氮矿化时空变化特征及其与土壤环境因子和酶活性之间的关系。结果表明：尕海湿地退化对土壤氮矿化过程有显著抑制作用,与未退化(0.143 mg kg^-1 d^-1)相比,轻度退化、中度退化、重度退化的土壤净氮矿化速率分别减小了0.018 mg kg^-1 d^-1、0.025 mg kg^-1 d^-1、0.020 mg kg^-1 d^-1;随着退化程度加剧,土壤净氨化速率逐渐减小或者不变,而净硝化速率却增大。随时间推移,各退化...     

1981—2018年新疆草地归一化植被指数时空特征及其对气候变化的响应

在新疆开展长时间序列的草地监测,分析草地生长的时空变化特征,有利于草地环境压力分析和草地生态健康预测。以NOAA-AVHRR NDVI为数据源,采用最大值合成、一元回归分析与相关性分析,分别在年际尺度和多个空间尺度(全疆、南北疆与各地区及其11种草地类型)上探讨了1981—2018年新疆草地归一化植被指数(NDVI)时空特征及其对气温、降水的响应。结果表明：(1)1981—2018年,新疆草地NDVI多年均值0.326,变化范围0.259—0.386,具有轻微年际波动特征;(2)北疆、南疆草地NDVI均表现为轻微增加趋势;全疆占草地总面积41%的区域NDVI呈显著增加趋势,9%为显著减少区域,北疆草地NDVI显著增加的面积是南疆的1.7倍;(3)由于垂直地带性及区域差异,新疆草地NDVI由山区向盆地的荒漠降低;北疆草地NDVI是南疆1.4倍,总体上北疆各地区草地NDVI高于南疆各地区;(4)草地类型植被NDVI对降水的显著响应高于气温,其中温性荒漠类、温性荒漠草原类与温性草原类草地NDVI对降水变化的响应明显高于其余草地类型,降水对草地NDVI的影响更为显著,表明降水引起的地表水分变化...     

河滨湿地不同植物群落根系分布特征与土壤理化性状的关系——以黄河中游荥阳段为例

在黄河中游郑州荥阳段,选择了5种河滨湿地植物群落进行根系和土壤性状特征研究,以期阐明不同植物群落的根系分布规律与土壤性状的关系,为河滨湿地植物群落组成以及土壤质量恢复提供科学参考。结果表明(1)在0—40 cm土层,根生物量密度与根长密度的平均值均表现为：芦苇群落(Phragmites australis)和芦苇-狗牙根(Cynodon dactylon)群落均大于芦苇-拂子茅(Calamagrostis epigeios)-狗牙根群落、拂子茅-狗牙根群落、拂子茅-狗牙根-水莎草(Juncellus serotinus)群落。拂子茅-狗牙根、芦苇-拂子茅-狗牙根、拂子茅-水莎草-狗牙根三种植物群落类型下根生物量密度、根长密度在0—20 cm表层土壤较大,芦苇群落和芦苇-狗牙根群落的根生物量密度、根长密度在10—40 cm的土层较大。(2)黄河河滨湿地芦苇群落、芦苇-狗牙根群落的土壤以粉粒为主,拂子茅-狗牙根群落、芦苇-拂子茅-狗牙根群落、拂子茅-狗牙根-水莎草群落的土壤主要以砂粒为主。在0—40 cm土层,芦苇群落、芦苇-狗牙根群落的土壤含水率、土壤有机质、有效氮和有效磷含量均显著高于...