应用纤维素结合域标签构建谷氨酸棒状杆菌表达系统

为优化谷氨酸棒状杆菌表达系统的纯化工艺,合成里氏木霉的CBD基因,将其与谷氨酸棒状杆菌分泌表达载体pXMJ19-sp连接,构建以CBD为纯化标签的重组载体pXMJ 19-sp-CBD.在该载体中插入GFP基因并转化至谷氨酸棒状杆菌,可获得分泌表达融合蛋白GFP-CBD的重组菌.该菌经IPTG诱导后的发酵液在紫外灯下显示强烈的绿色荧光,重组蛋白的分泌表达量达200 mg/L.利用CBD标签对纤维素柱的可逆性吸附,可直接对谷氨酸棒状杆菌分泌到培养基中的重组蛋白进行纯化,从而简化工艺和降低成本,为工业化大生产奠定基础.     

酱香型白酒酿造酒醅中酵母菌多样性研究

为解析酱香型白酒酿造酒醅中酵母菌的菌群结构,获取酒醅中的主要酵母菌,采用高通量测序法分析酱香型白酒酒醅中酵母菌多样性及主要功能菌群,同时采用可培养分离方法获取酒醅中酵母菌活性菌株。从酱香型白酒下沙至五轮次酒醅中共检出59个属、129个种的酵母菌,分离得到酵母菌活性菌株41种,检测到的酵母菌种类与获得的酵母菌活菌在各香型白酒中最多。不同时期酒醅中的酵母菌种类和数量差异明显,其中下沙、造沙轮次以Pichia kudriavzevii为绝对优势酵母菌;一至五轮次随着轮次的递增,酒醅中优势酵母菌的种类增多,其中主要的优势酵母菌有Pichia kudriavzevii、Pichia manshurica、Zygosaccharomyces bailii、Saccharomyces cerevisiae、Candida apicola。酱香型白酒酒醅中蕴藏着极其丰富的酵母菌资源,对酵母菌菌群结构的解析有助于科学地认识酱香型白酒酿造过程中产酒与风味代谢机理,为发酵过程的调控提供一定依据。     

鲍姆纤孔菌（桑黄）不同方式发酵的菌丝体生物活性比较

利用液体发酵、木屑固体发酵和米饭固体发酵3种方式培养鲍姆纤孔菌（桑黄）菌丝体,对菌丝体醇提物的体外抗氧化、抗肿瘤和抗衰老生物活性进行了研究。结果表明,木屑固体发酵、液体发酵和米饭固体发酵的菌丝体醇提物清除H2O2自由基的IC50值分别为78.28±0.32、27.73±0.57和7.84±0.37;米饭培养的桑黄菌丝体醇提物在低浓度500μg/mL下对超氧阴离子自由基清除作用到达80%,在相同的浓度下对DPPH自由基清除率也明显高于其他两种方法,表现出较高的抗氧化活性。木屑以及米饭培养方法得到的菌丝体对PC12神经细胞损伤修复均有较好的效果,液体培养的桑黄菌丝体表现的修复作用较低;液体发酵培养的菌丝体醇提物浓度在100μg/mL时,对肿瘤细胞HepG2的抑制率达70%,高于其他两种培养方法的抑制作用。     

中国西藏种子植物区系新资料

本文报道了采自西藏喜马拉雅南坡的8个中国种子植物新记录种以及1个西藏新记录属。前者分别是吉隆牛奶菜(Marsdenia roylei)、塔基棕榈(Trachycarpus takil)、喀西蜂斗草(Sonerila khasiana)、旋花锡生藤(Cissampelos convolvulacea)、吉隆角盘兰(Herminium edgeworthii)、尼泊尔西番莲(Passiflora napalensis)、椭穗姜花(Hedychium ellipticum)和藏南象牙参(Roscoea brandisii); 1个西藏新记录属为箭药藤属(Belostemma) (箭药藤 Belostemma hirsutum)。凭证标本存放于中国科学院西双版纳热带植物园标本馆(HITBC)和西藏自治区高原生物研究所标本室(XZ)。     

The role of transferrin in natural killer cell and IL-2-induced cytotoxic cell function

The growth factor transferrin (Tf) enhanced natural killer (NK) cell cytotoxicity. This enhancement was due to direct effects on NK cell function, and Tf treatment of the K562 target cell had no effect on their sensitivity. NK cells were highly enriched in the low-density large granular lymphocyte population (LGL) by Percoll gradient centrifugation. Despite the direct effect of Tf on NK cells, the number of cells expressing receptors for Tf (TfR) in NK-enriched LGL was the same as the NK-cell-depleted high-density small lymphocyte population (SL). All populations, tested without stimulation, had very few TfR+ cells. Interleukin 2 (IL-2) could induce very high NK-like activity in the LGL but not in SL. Similarly, only LGL could be induced by IL-2 to express TfR. In serum-free cultures, only limited NK-like activity could be developed which was greatly enhanced by supplementing with Tf in the cultures. The importance of Tf in NK-like development was confirmed by modulating the expression of TfR in IL-2 containing cultures with mouse monoclonal antibody OKT9 specific for TfR. OKT9 totally abrogated the induction of cytotoxic activity by IL-2 against K562 and NK-resistant target. OKT9 inhibited the induction of cytotoxicity in both lymphocytes containing active NK cells and in those predepleted of active NK cells, indicating that the development of NK-like activity from both precursor populations requires Tf. The inhibition by OKT9 was only during the induction phase. The same antibody had no effect on the cytotoxicity of fresh NK cells or the mature IL-2-induced NK-like cells. Our data therefore do not support the hypothesis of TfR as the NK recognition structure. Instead, these results indicate that Tf is important for the development of NK and NK-like activities.     

北京地区红隼的迁徙路线和活动区域

红隼(Falco tinnunculus)被列为国家二级重点保护野生动物,是能同时适应农村和城市环境的小型猛禽,对维持城市生态系统稳定具有重要意义。2022年4月至7月,为在北京救助的7只红隼佩戴了卫星追踪器,追踪其活动轨迹,依据追踪的动物活动位点数据,采用净平方位移-时间曲线依次对各红隼的迁徙模式进行了判别,深入分析了迁徙红隼的迁徙时间、距离和路线等,并采用核心密度法分别计算了迁徙及留居型红隼95%及50%活动区面积。研究结果表明,在北京地区红隼的迁徙模式为部分迁徙,追踪的7只红隼个体(N01 ~ N07)中,4只为留鸟,1只为迁徙鸟,2只居留类型无法准确判断。N01为迁徙红隼,其度夏地和越冬地分别在内蒙古锡林郭勒盟和河北廊坊,此红隼秋季迁徙速度明显高于春季,其春季迁徙距离551 km,历时25 d,平均迁徙速度为22 km/d,而秋季迁徙距离412 km,历时2 d,平均迁徙速度为203 km/d,河北承德滦平县是其春季迁徙的重要中途停歇地。不同红隼个体间95%及50%活动区面积均存在较大差异,迁徙红隼N01 95%、50%活动区面积在度夏区分别为93.10 km²、17.50 km²,在越冬区分别为7.03 km²、0.99 km²;留居型红隼95%、50%活动区面积均值分别为1 165.34 km²、178.71 km²(n = 4),其中最大95%、50%活动区面积分别为4 320.26 km²(N02)、648.22 km²(N02),最小95%、50%活动区面积分别为2.80 km²(N03)、0.29 km²(N03)。本研究揭示了北京地区红隼的迁徙模式、迁徙路线、重要停歇地及活动区状况,为红隼的针对性保护和管理提供了科学依据。     

湖南沅江鼎城段河蚬的性别组成与精子形态特征

本研究以湖南沅江鼎城段河蚬(Corbicula fluminea)为研究对象,在进一步验证该群体性别组成的基础上,分析壳长组成与性别比例之间关系,探讨该群体雄性、雌性和雌雄同体之间关系;同时以单鞭毛精子为参照,分析比较双鞭毛精子的形态特征,以期为我国河蚬的性别发生及生殖适应策略研究提供基础资料。结果显示：沅江鼎城段河蚬(n = 770)雄性、雌性和雌雄同体最小性成熟壳长分别为2.92 mm、5.66 mm和5.30 mm。697只性成熟河蚬中雄性、雌性和雌雄同体的比例近似1︰1︰6。雌雄同体的平均壳长[(22.55 ± 0.33)mm,n = 517]显著大于雄性[(20.44 ± 1.03)mm,n = 95]和雌性[(19.79 ± 0.99)mm,n = 85](P < 0.05),但雄性与雌性的平均壳长之间差异不显著(P > 0.05)。河蚬可以产生单鞭毛和双鞭毛两种类型的精子,单鞭毛精子头长范围4.93 ~ 21.79 μm,平均值(14.27 ± 0.82)μm(n = 30),双鞭毛精子头长范围10.29 ~ 22.04 μm,平均值(15.62 ± 0.62)μm(n = 26)。单、双鞭毛精子头长差异不显著(P > 0.05)。双鞭毛精子(n = 26)长尾的平均长度[(38.07 ± 1.44)μm]显著大于其短尾[(31.08 ± 1.60)μm]和单鞭毛精子(n = 30)尾部长度[(30.15 ± 1.75)μm](P < 0.01),但其短尾与单鞭毛精子的尾部平均长度之间无显著性差异(P > 0.05)。结果表明：湖南沅江鼎城段河蚬为雄性先熟,且可能存在雄性和(或)雌性向雌雄同体转换现象。河蚬具有单鞭毛型和双鞭毛型2种类型的精子,且双鞭毛精子的2个尾部不等长。同域共存河蚬的单鞭毛与双鞭毛精子在运动及受精能力方面的差异值得深入研究。     

Lectin analysis of common glycoproteins detected on the surface of continuous microvascular endothelium in situ and in culture: identification of sialoglycoproteins

For many years, molecular interactions with vascular endothelium have been studied in vitro on cultured endothelial cells. Yet, it is clear that the different environmental conditions in vivo vs. in vitro may cause phenotypic drift and altered expression of cell surface molecules. In this study, we identify several endothelial surface proteins of similar apparent molecular mass by radioiodination of cultured microvascular cells and by intravascular radioiodination of rat heart endothelium in situ. The radioiodinated surface polypeptides detected by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) (followed by autoradiography) were subjected to lectin affinity chromatography in order to provide an additional screen for identifying common surface glycoproteins and a means for partial characterization of their glycans. With a battery of 18 lectins, seven major (gp140, gp120, gp100, gp85, gp75, gp60, gp47) and 6 minor (gp330, gp300, gp180, gp160, gp150, gp42) glycoproteins were identified on the cultured cells each with a different lectin binding profile. The lectin binding profiles of many endothelial glycoproteins in situ were similar to those of their counterparts in culture. A common set of seven major glycoproteins with the same apparent molecular masses was found in situ as well as in vitro. These common glycoproteins were characterized further using both sialidase digestion and sequential lectin affinity chromatography of cell lysates. Most of the glycoproteins appear to have both complex-type N-linked and O-linked glycans except for gp60 with only O-linked glycans, gp47 with only complex N-linked sugars, and gp42 with only simple N-linked sugars. A subset of sialoglycoproteins (gp140, gp120, gp100, gp60, gp47) was identified. One of them, gp120, is podocalyxin based on immunoprecipitation with specific antiserum and another one, gp60, is a recently identified albumin binding protein on the surface of cultured microvascular endothelial cells. This study shows that gp60 is indeed present on the surface of endothelium in situ and that it is a sialoglycoprotein with typical O-linked glycans. It is apparent that the continuous type of microvascular endothelium can indeed express in culture and in situ a common set of major glycoproteins.     

三种海水生态养殖池塘细菌数量特征及其与环境因子的相关性

为阐明不同生态养殖池塘浮游细菌数量变化规律及其与环境因子间的相关性,以期为多元立体综合养殖模式中的水质安全与生物疾病防控提供科学依据。利用荧光显微镜细菌计数法(Acridine Orange Direct Counting),研究了“参-虾”“蟹-蛏”和“蜇-蛏”三种海水生态养殖池塘的细菌数量特征,使用典范对应分析(Canonical Correspondence Analysis, CCA)探讨细菌数量与环境因子的相互关系。结果表明,“参-虾”“蟹-蛏”和“蜇-蛏”池塘的细菌平均密度分别为(0.49～3.32)×10⁵ cell/mL、(0.52～2.98)×10⁵ cell/mL和(0.47～2.55)×10⁵ cell/mL,细菌数量有明显的时间差异,夏季密度大于春秋季。三种海水生态养殖池塘的水温、溶解氧和pH变化均不明显,且相差不大,而硝态氮均发生显著变化;其中“蟹-蛏”和“蜇-蛏”池塘的硝态氮、亚硝态氮、活性磷、总氮和总磷均高于“参-虾”池塘,透明度、叶绿素a和化学耗氧量则均低于“参-虾”池塘。典范对应分析(C...