Qualitative and quantitative analysis of the adult Drosophila melanogaster proteome

Drosophila melanogaster is one of the most widely used model organisms in life sciences. Mapping its proteome is of great significance for understanding the biological characteristics and tissue functions of this species. However, the comprehensive coverage of its proteome remains a challenge. Here, we describe a high‐coverage analysis of whole fly through a 1D gel electrophoresis and LC‐MS/MS approach. By combining the datasets of two types of SDS‐PAGE and two kinds of tagmata, the high‐coverage analysis resulted in the identification of 5262 genes, which correspond to 38.23% of the entire coding genes. Moreover, we found that the fly head and body have different molecular weight distributions of their proteomes when the proteins were resolved with SDS‐PAGE and image analysis of the stained gel. This phenomenon was further confirmed by both label‐free and isobaric tags for relative and absolute quantitation‐based quantitative approaches. The consistent results of the two different quantitation methods also demonstrated the stability and accuracy of the LC‐MS/MS platform. The MS proteomics data have been deposited to the ProteomeXchange with identifiers PXD000454 and PXD000455 ( http://proteomecentral.proteomexchange.org/dataset/PXD000454 ; ( http://proteomecentral.proteomexchange.org/dataset/PXD000455 ).     

罗望子中葡聚木糖的结构与功能初探

乔木植物罗望子种子中富含木聚葡糖,是一种理想的膳食纤维来源。本文对该种子中所含的可溶性纤维进行了纤维素酶水解－ＨＰＬＣ分析和甲基化键型分析,证明其结构与在细胞壁中木聚葡糖基本相同。小白鼠动物实验初步确定了其促进肠蠕动的作用。     

Estimation of amino acid pairs sensitive to variants in human phenylalanine hydroxylase protein by means of a random approach

In this data-based theoretical analysis, we use a random approach to estimate amino acid pairs in human phenylalanine 4-hydroxylase (PAH) protein in order to determine which amino acid pairs are more sensitive to 187 variants in human PAH protein. The rationale of this study is based on our hypothesis and previous findings that the harmful variants are more likely to occur at randomly unpredictable amino acid pairs rather than at randomly predictable pairs. This is reasonable to argue as randomly predictable amino acid pairs are less likely to be deliberately evolved, whereas randomly unpredictable amino acid pairs are probably deliberately evolved in connection with protein function. 94.12% of 187 variants occurred at randomly unpredictable amino acid pairs, which accounted for 71.84% of 451 amino acid pairs in human PAH protein. The chance of a variant occurring is five times higher in randomly unpredictable amino acid pairs than in predictable pairs. Thus, randomly unpredictable amino acid pairs are more sensitive to variance in human PAH protein. The results also suggest that the human PAH protein has a natural tendency towards variants.     

光学显微镜技术在活细胞单分子检测中的应用

单分子检测是一门以高度的时间以及空间分辨率研究生物单分子的技术。近来,科学技术的探索发展使我们可以观察、检测甚至操纵单个分子并且研究它们的构象变化和动力学行为。这一发展使得以前被传统系综研究体系平均化所隐藏的新信息被揭示出来。单分子检测技术的发展已经揭开了生命科学研究的新篇章。在本文中,我们将介绍有关活细胞中单分子检测技术的发展以及活细胞内单分子检测的现状。     

mMR-1基因的克隆和在毕赤酵母中分泌表达

hMR-1为本实验室首次克隆发现的一个人类新基因 ,是一种和三种肌肉收缩蛋白及多种细胞信号蛋白具有相互作用的膜蛋白。经与鼠基因组数据库进行同源分析后设计合成引物 ,利用RT-PCR技术从小鼠C57BL 6J脾脏T淋巴细胞总RNA中反转录得到鼠源MR-1基因 (mMR-1) ,提交GenBank ,收录号 (AY2 99972 )。序列分析证明其与人源MR-1基因 (hMR-1)同源性为 90.1%。构建表达载体pPIC9 mMR-1电转化PichiapastorisGS115 ,筛选得到整合分泌表达mMR-1蛋白的重组酵母菌株。表达目的蛋白分子量 25kD ,诱导 5d时产量达到 50mg/L ,通过Westernblot验证了表达产物的正确性。本研究为进一步研究新基因MR-1的生物学功能奠定了基础。     

SSH 法结合定量 PCR 技术研究双肌臀猪 肌肉组织的差异表达基因

利用抑制性消减杂交技术 (suppression subtractive hybridization , SSH) 成功地构建了双肌臀与非双肌臀大白猪肌肉组织差异表达的消减 cDNA 文库,获得有效克隆 686 个. 对整个文库测序分析,共获得 587 条有效序列. 利用 BLAST 在线软件与 GenBank、 GenBank EST 和 Tigr Porcine EST 等数据库进行同源序列比较,发现其中有 11 个未知新序列,可能代表“双肌臀”大白猪肌肉组织特异表达的新基因. 对文库中所包含的兰尼啶受体基因 (RYR1)、钙依赖性蛋白激酶 基因 (CAMK2)、人类胰岛素生长因子结合蛋白 -7 基因 (IGFBP7),以及 695号、 882号和480号3个新基因进行了实时荧光定量 PCR 检测,结果显示： RYR1、 CAMK2 及 IGFBP7 基因在双肌臀猪肌肉组织 RNA 池中的表达量,分别为对照的非双肌臀猪肌肉组织 RNA 池中表达量的 1.87、 1.90 和 1.85 倍; 695 号、 882 号和 480 号 3 个新的 ESTs 在双肌臀猪肌肉组织 RNA 池中的表达量为非双肌臀猪肌肉组织 RNA 池中表达量的 1.48、 1.44 和 1.78 倍. 这提示我们,上述基因的上调表达很可能与猪双肌臀性状的发生有密切的关系,可以作为研究该性状的候选基因.     

鲸类分子系统学研究进展

综述了近年来分子生物学标记技术在鲸类系统学研究中的进展。分子生物学证据支持鲸目与有蹄类之间有较近的亲缘关系,并支持鲸类的单系起源,但鲸类不同类群(须鲸类、抹香鲸类及不包括抹香鲸类的齿鲸类)之间的系统发生关系仍存在争议。抹香鲸类到底与须鲸类还是与其它齿鲸类有更近的亲缘关系,不同的分子生物学家所得到的结果并不一致。此外,分子生物学技术还被用于解决须鲸亚目和齿鲸亚目内科间以及科内种间的系统发生关系,特别是齿鲸亚目的海豚科、鼠豚科和淡水豚类。通过分子标记技术来研究鲸类种下的遗传结构是鲸类分子系统学研究中的一个新热点,使用的标记主要是mtDNA控制区、核DNA微卫星和主要组织相容性复合体(major histo-compatibilitv complex,MHC)等。     

序贯式止血法用于腹腔镜卵巢囊肿剥除术的临床研究

目的：探讨腹腔镜卵巢囊肿剥除术中采用序贯式止血法对卵巢功能的影响。方法：将100例行腹腔镜卵巢囊肿剥除术患者按照抽签法随机地均分为对照组与观察组,对照组给予传统双极电凝治疗,观察组采取序贯式止血法,靠近卵巢门的髓质出血镜下缝合,远离卵巢门行皮质渗血电凝处理。比较两组患者手术前后雌二醇（Estradiol,E2）、促卵泡激素（Follicle-stimulating hormonc,FSH）、促黄体生成素（Luteotropic hormone,LH）及围绝经期综合征发生情况。结果：对照组术前与术后3个月及术后6个月E2、FSH及LH差异均具有统计学意义,观察组手术前后激素水平差异无统计学意义,且观察组术后与对照组术后上述指标差异均具有统计学意义;对照组患者术后月经紊乱、植物神经功能紊乱的发生率分别为14．00％与12．00％,观察组为2．00％及0％,两组差异具有显著的统计学意义。结论：序贯式止血法应用于腹腔镜卵巢囊肿剥除术中影响卵巢功能,效果显著,值得在临床上加以推广并应用。     

Different types of nitrogen deposition show variable effects on the soil carbon cycle process of temperate forests

Nitrogen (N) deposition significantly affects the soil carbon (C) cycle process of forests. However, the influence of different types of N on it still remained unclear. In this work, ammonium nitrate was selected as an inorganic N (IN) source, while urea and glycine were chosen as organic N (ON) sources. Different ratios of IN to ON (1 : 4, 2 : 3, 3 : 2, 4 : 1, and 5 : 0) were mixed with equal total amounts and then used to fertilize temperate forest soils for 2 years. Results showed that IN deposition inhibited soil C cycle processes, such as soil respiration, soil organic C decomposition, and enzymatic activities, and induced the accumulation of recalcitrant organic C. By contrast, ON deposition promoted these processes. Addition of ON also resulted in accelerated transformation of recalcitrant compounds into labile compounds and increased CO₂ efflux. Meanwhile, greater ON deposition may convert C sequestration in forest soils into C source. These results indicated the importance of the IN to ON ratio in controlling the soil C cycle, which can consequently change the ecological effect of N deposition.     

TILLING技术在功能基因组学中的应用

TILLING(定向诱导基因组局部突变)技术是近年发展起来的一种高通量筛选化学诱变的点突变的技术,它利用专一识别点突变的核酸酶结合PCR来检测单核苷酸多态性(SNP)。TILLING技术起源于植物基因组研究,逐渐扩展到动物及人类功能基因组学的研究中。无论是筛选突变体还是研究特定基因的重要性,TILLING都具有高通量、自动化的优势。随着此项技术应用范围的扩展,从诱变剂和内切酶的选择到具体的操作方式,以及结果的识别和统计方法,都有了不少改进。在其他相关学科不断发展的大环境下,TILLING技术也在不断发展,其在功能基因组学研究中的作用也会更显著。