远交系小鼠胚胎干细胞系的建立及嵌合鼠的获得

ＥＳ细胞（ＥｍｂｒｙｏｎｉｃＳｔｅｍＣｅｌｌｓ）是来源于小鼠早期胚胎的多潜能干细胞,它可以在体外大量培养。并以单细胞的形式注射到早期胚胎里,发育为嵌合体。到目前为止,通常使用的１２９小鼠品系是来源于近交系（ｉｎｂｒｅｄ）小鼠的胚胎．与之相比,远交系小鼠应当具有较强的生命力和抗病能力。曾有人报道过建成了远交系小鼠胚胎干细胞系,但是尚没有见到获得嵌合鼠的报道。有人甚至认为：由于不同品系小鼠所具有的遗传背景不同,有的小鼠不能建成ＥＳ细胞系。最近,本实验室在这方面做了有益的探索,成功地建成了远交系小鼠胚胎干细胞系,并在这里报导首例用远交系小鼠胚胎干细胞系培育成功嵌合体小鼠。采用源于Ｓｗｉｓｓ小鼠远交群的昆明（ＫＭ）品系小鼠囊胚建成了三个小鼠胚胎干细胞系（ＫＥ１．ＫＥ２．ＫＥ５）。核型正常率均达到７０％以上。自第八代起分批冻存,复苏后,培养至第１２代,消化成单细胞,通过囊胚显微注射,将其注射到６１５品系小鼠胚胎。在幸存的幼鼠中获得了一只来源于ＫＥ１细胞的嵌合鼠（Ｔａｂｌｅ１）．其毛色表现为受体鼠（６１５）的白色中嵌合有供体鼠（ＫＭ）的黑褐色（ＰｌａｔｅＩ－Ａ）．嵌合鼠与受体鼠的杂交后代鼠中仍然出现了受体鼠的毛色类型（     

Targeting foreign proteins to human immunodeficiency virus particles via fusion with Vpr and Vpx.

The human immunodeficiency virus type 1 (HIV-1) and HIV-2 Vpr and Vpx proteins are packaged into virions through virus type-specific interactions with the Gag polyprotein precursor. To examine whether HIV-1 Vpr (Vpr1) and HIV-2 Vpx (Vpx2) could be used to target foreign proteins to the HIV particle, their open reading frames were fused in frame with genes encoding the bacterial staphylococcal nuclease (SN), an enzymatically inactive mutant of SN (SN*), and chloramphenicol acetyltransferase (CAT). Transient expression in a T7-based vaccinia virus system demonstrated the synthesis of appropriately sized Vpr1-SN/SN* and Vpx2-SN/SN* fusion proteins which, when coexpressed with their cognate p55Gag protein, were efficiently incorporated into virus-like particles. Packaging of the fusion proteins was dependent on virus type-specific determinants, as previously seen with wild-type Vpr and Vpx proteins. Particle-associated Vpr1-SN and Vpx2-SN fusion proteins were enzymatically active, as determined by in vitro digestion of lambda phage DNA. To determine whether functional Vpr1 and Vpx2 fusion proteins could be targeted to HIV particles, the gene fusions were cloned into an HIV-2 long terminal repeat/Rev response element-regulated expression vector and cotransfected with wild-type HIV-1 and HIV-2 proviruses. Western blot (immunoblot) analysis of sucrose gradient-purified virions revealed that both Vpr1 and Vpx2 fusion proteins were efficiently packaged regardless of whether SN, SN*, or CAT was used as the C-terminal fusion partner. Moreover, the fusion proteins remained enzymatically active and were packaged in the presence of wild-type Vpr and Vpx proteins. Interestingly, virions also contained smaller proteins that reacted with antibodies specific for the accessory proteins as well as SN and CAT fusion partners. Since similar proteins were absent from Gag-derived virus-like particles and from virions propagated in the presence of an HIV protease inhibitor, they must represent cleavage products produced by the viral protease. Taken together, these results demonstrate that Vpr and Vpx can be used to target functional proteins, including potentially deleterious enzymes, to the human or simian immunodeficiency virus particle. These properties may be exploitable for studies of HIV particle assembly and maturation and for the development of novel antiviral strategies.     

籼稻的抗冷性与亲本的关系

四个籼稻（Ｏｒｙｚａ ｓａｔｉｖａ Ｌ．）品种幼苗经１℃黑暗或光照２５０ μｍ ｏｌ·ｍ － ２·ｓ－ １处理后,抗冷的“桂山矮选３”比不抗冷的“青华６ 号”幼苗存活率高,其子代是以“桂山矮选３”为母本的比“青华６ 号”为母本的存活率较高。抽穗期剑叶经光照低温处理１２、２４ 和３６ ｈ 后,光合作用是“桂山矮选３”和以“桂山矮选３”为母本的子代比“青华６ 号”和以“青华６ 号”为母本的子代下降较少。呼吸作用是前者比后者在处理１２ ｈ 时有明显升高现象。荧光参数Ｆｖ／Ｆｏ和Ｆｖ／Ｆｍ比值在处理２４ ｈ 时前者比后者下降明显,但在常温下恢复则是前者比后者明显较快。自然低温（寒露风）对叶绿素荧光的影响亦有相似的规律。对水稻后代的抗冷性倾向于母本进行了讨论     

菜豆热激蛋白在生物膜上的定位

选用菜豆 Phaseolus vulgris L. 下胚轴 ,运用35S- Met标记放射自显影和二维电泳技术 ,研究热激蛋白 HSPs 的表达和在生物膜组分中的定位 .实验结果表明 ,盐溶蛋白中主要HSPs为 70 k D HSPs和小分子量 HSPs,而小分子量组 HSPs大量富集在质膜和液泡膜组分中 .     

棒状类细菌电击转化中多种条件对转化效率的影响

以质粒PXZl0145电击转化不同棒状类细菌菌株,研究了影响电击转化效率的诸个因素,在含4%甘氨酸的培养基中生长至对数前期的菌体最适用于电击转化,当以同源DNA进行电击转化时,1．μgDNA中转化效率最高可达到8×1O⁶转化子,但用异源DNA时,转化效率要比前者低10²～10³倍。     

Evidence forCis- andTrans-acting element coevolution of the 2-μm circle genome inSaccharomyces cerevisiae

Summary We compared the DNA sequence of the yeas 2-μm plasmidcis-actingSTB andtrans-actingREP1 partition loci of laboratory haploid and industrial amphiploid strains. Several industrial strains had a uniqueSTB sequence (type 1) sharing only 70% homology with laboratorySTB (type 2). Type 1 plasmids had a REP1 protein with 6–10% amino acid substitutions when compared to REP1 of type 2 plasmids. All 2-μm variants that shared a similarSTB consensus sequence exhibited a high degree ofREP1 nucleotide and amino acid sequence conservation. These observations suggest molecular coevolution oftrans-acting elements with cognate target DNA structure. Based on DNA sequencing and Southern hybridization analyses, we classified 2-μm variants into two main evolutionary lineages that differ atSTB as well asREP1 loci. The role of molecular coevolution in yeast intra- and interspecies plasmid evolution was discussed.     

Spatial and temporal patterns of intracellular calcium in colonic smooth muscle

Summary Intracellular calcium [Ca²⁺] _i measurements in cell suspension of gastrointestinal myocytes have suggested a single [Ca²⁺] _i transient followed by a steady-state increase as the characteristic [Ca²⁺] _i response of these cells. In the present study, we used digital video imaging techniques in freshly dispersed myocytes from the rabbit colon, to characterize the spatiotemporal pattern of the [Ca²⁺] _i signal in single cells. The distribution of [Ca²⁺] _i in resting and stimulated cells was nonhomogeneous, with gradients of high [Ca²⁺] _i present in the subplasmalemmal space and in one cell pole. [Ca²⁺] _i gradients within these regions were not constant but showed temporal changes in the form of [Ca²⁺] _i oscillations and spatial changes in the form of [Ca²⁺] _i waves. [Ca²⁺] _i oscillations in unstimulated cells (n = 60) were independent of extracellular [Ca²⁺] and had a mean frequency of 12.6 +1.1 oscillations per min. The baseline [Ca²⁺], was 171 ± 13 nm and the mean oscillation amplitude was 194 ± 12 nm. Generation of [Ca²⁺] _i waves was also independent of influx of extracellular Ca²⁺. [Ca²⁺] _i waves originated in one cell pole and were visualized as propagation mostly along the subplasmalemmal space or occasionally throughout the cytoplasm. The mean velocity was 23 +3 m per sec (n = 6). Increases of [Ca²⁺] _i induced by different agonists were encoded into changes of baseline [Ca²⁺] _i and the amplitude of oscillations, but not into their frequency. The observed spatiotemporal pattern of [Ca²⁺] _i regulation may be the underlying mechanism for slow wave generation and propagation in this tissue. These findings are consistent with a [Ca²⁺] _i regulation whereby cell regulators modulate the spatiotemporal pattern of intracellularly generated [Ca²⁺] _i oscillations.The authors thank Debbie Anderson for excellent technical assistance with the electron microscopy and Dr. M. Regoli for providing the NK-1 agonist [Sar⁹,Met(O₂)¹¹]-SP. This work was supported by National Institutes of Health Grants DK 40919 and DK 40675 and Veterans Administration Grant SMI.     

Distribution and behaviour of Acetes sibogae Hansen (Decapoda, Crustacea) in an estuary in relation to tidal and diel environmental changes

Quantitative samples of Acetes sibogae were collected at 2 hintervals for 48 h at three sites across the axis of a tidalestuary to examine their distribution within the water bodyover tidal and diel cycles, and to assess the role of behaviourin maintaining population distribution in estuarine/coastalwaters in relation to selected environmental factors. Watertemperature, salinity, tidal height and light intensity wereconcurrently measured. Distribution of the shrimp across theestuary was uniform and consistent between daylight or darkperiods, and among flood or ebb tides. Changes of A.sibogaeabundance were related to light and tidal cycles at each sitewith higher catches in dark periods and during flood tides.Acetes sibogae also exhibited both nocturnal and tidal verticalmovements in the water body, with greater numbers being onlyfound near-surface rather than near-bottom during flood tidesand at night. No significant differences in the distributionof size groups were found between any sampled levels of anysite. Acetes sibogae was highly aggregated in the water body.It is suggested that aggregating behaviour and tidal and nocturnalvertical movements act to facilitate population maintenancein estuarine/coastal waters.     

中国虫草一新记录种

【目的】对一个寄生鳞翅目幼虫的虫草标本Dxhir140901进行分类鉴定。【方法】采用形态学比较和基于ITS1-5.8S-ITS2rDNA的系统发育与进化网络分析进行鉴定。【结果】形态学观察：标本的分离菌株形态显示其为典型的被毛孢属真菌,具有两型产孢结构：A型产孢细胞柱状,(1.8?6.3) μm×1.8 μm;B型产孢细胞锥形,基部柱状,向上逐渐变细无明显颈部,基部宽3?3.8 μm,长21?63 μm,颈部宽1.8?2.0 μm,菌丝末端可直接形成产孢细胞;孢子橘瓣形或卵形,(8.1?10.8) μm×(2.7?5.4) μm,具粘液,黏液层厚1.8?2.7 μm。系统发育分析结果显示该菌株与巨针线形虫草Ophiocordyceps macroacicularis聚为一支,支持率为98%,进化网络分析也支持上述结果。【结论】通过与O. macroacicularis的形态比较和分子系统学分析结果,Dxhir140901及其分离株Gzuifr-hir140901为巨针线形虫草Ophiocordyceps macroacicularis S. Ban, T. Sakane & Nakagiri的无性阶段,该种为中国新记录种。