禽病原性大肠杆菌1型菌毛的分离与鉴定

以旋涡混合法使禽病原性大肠杆菌分离株566、1794和TK3菌毛脱落,经硫酸铵沉淀、透析后进行蔗糖密度梯度离心,收集密度为110至115g/cm3的蛋白带,经SDSPAGE测定,3株菌菌毛蛋白的分子量分别在175、170和170kD;提纯菌毛保留了甘露糖敏感性凝集豚鼠红细胞的能力,证明它们为1型菌毛;从1794株提取的1型菌毛免疫BALB/C小鼠产生的高免血清在Western blot中与3个菌株的相应菌毛蛋白均呈阳性反应。上述结果表明,受检的3株禽病原性大肠杆菌均表达了1型菌毛,其分子量在175～170kD之间,3个菌株的1型菌毛间具有较强的抗原相关性。     

多能干细胞诱导分化为肾脏类器官的研究进展与挑战

用于分化为多种类型细胞的多能干细胞(PSC)体外培养技术已被广泛应用于生物学领域中.由PSC分化而来的肾脏类器官可基本还原生物体内肾脏的组织结构和部分功能,在肾脏疾病模型研究和药物筛选中有重要作用,继续改善肾脏类器官的结构、功能和成熟度将会对肾脏再生治疗提供极大的帮助.研究肾脏类器官的重点在于体外准确模拟体内肾脏的发育...     

Generation of virus‐resistant potato plants by RNA genome targeting

CRISPR/Cas systems provide bacteria and archaea with molecular immunity against invading phages and foreign plasmids. The class 2 type VI CRISPR/Cas effector Cas13a is an RNA‐targeting CRISPR effector that provides protection against RNA phages. Here we report the repurposing of CRISPR/Cas13a to protect potato plants from a eukaryotic virus, Potato virus Y (PVY). Transgenic potato lines expressing Cas13a/sgRNA (small guide RNA) constructs showed suppressed PVY accumulation and disease symptoms. The levels of viral resistance correlated with the expression levels of the Cas13a/sgRNA construct in the plants. Our data further demonstrate that appropriately designed sgRNAs can specifically interfere with multiple PVY strains, while having no effect on unrelated viruses such as PVA or Potato virus S. Our findings provide a novel and highly efficient strategy for engineering crops with resistances to viral diseases.     

Phycobilisome from Anabaena Variabilis Kütz. and its Model Conjugates

The model conjugates phycocyanin-allophycocyanin (C-PC-APC) and phycoerythrocyanin-phycocyanin-allophycocyanin (PEC-C-PC-APC) were synthesized by using a heterobifunctional coupling reagent N-succinimidyl-3-(2-pyridyldithio)propionate. The rod-core complex (αβ)₆ ^PCL_RC ²⁷(αβ)₃ ^APCL_C ^8.9 and phycobilisomes were separated from Anabaena variabilis. Energy transfer features for the conjugates and the complexes were compared. The absorption and fluorescence emission spectra indicated that the linker-peptides mediate interaction of phycobiliproteins and prompt energy transfer. The energy transfer in the conjugates was detected by fluorescence emission spectra and confirmed by the addition of dithiothreitol. The conjugates may be used as models for studying the energy transfer mechanism in phycobilisomes. This revised version was published online in September 2006 with corrections to the Cover Date.     

A novel thermostable phytase from the fungus Aspergillus aculeatus RCEF 4894: gene cloning and expression in Pichia pastoris

A novel gene of thermostable phytase, phyA, was isolated by polymerase chain reaction (PCR) techniques from Aspergillus aculeatus RCEF 4894. The full-length phyA gene comprises 1,404 bp and encodes 467 amino-acid residues, including a 19-residue putative N-terminal signal peptide. The phytase of A. aculeatus was a novel addition to the histidine-acid phosphatase family, as evidenced by both the conserved motifs RHGXRXP and HD in the amino-acid sequence, and 3D structure models. The recombinant phytase was overexpressed in Pichia pastoris, and its specific activity reached 3,000 U mL⁻¹ at the optimum pH of 5.5. This recombinant, thermostable phytase was able to withstand temperatures of up to 90 °C for 10 min, with a loss of only 13.9% of initial enzymatic activity, and showed high activity with phytic-acid sodium salt at a pH range of 2.5–6.5. The broad pH optima and high thermostability of the phytase makes it a promising candidate for feed-pelleting applications.     

关于提高血气分析仪测量精度的探讨

血气分析已广泛用于昏迷、休克、严重外伤的危急病人的血液酸碱度、氧分压、二氧化碳分压的测量。测量数据的准确性至关重要。本文对与测量精度有关的几个环节如计量检定、质控与定标、样本采集与测量、日常维护与故障处理进行探讨,提出提高血气分析精度的解决方案。     

Human endothelial cell growth and phenotypic expression on three dimensional poly(lactide-co-glycolide) sintered microsphere scaffolds for bone tissue engineering

Bone tissue engineering offers promising alternatives to repair and restore tissues. Our laboratory has employed poly(lactide-co-glycolide) PLAGA microspheres to develop a three dimensional (3-D) porous bioresorbable scaffold with a biomimetic pore structure. Osseous healing and integration with the surrounding tissue depends in part on new blood vessel formation within the porous structure. Since endothelial cells play a key role in angiogenesis (formation of new blood vessels from pre-existing vasculature), the purpose of this study was to better understand human endothelial cell attachment, viability, growth, and phenotypic expression on sintered PLAGA microsphere scaffold. Scanning electron microscopy (SEM) examination showed cells attaching to the surface of microspheres and bridging the pores between the microspheres. Cell proliferation studies indicated that cell number increased during early stages and reached a plateau between days 10 and 14. Immunofluorescent staining for actin showed that cells were proliferating three dimensionally through the scaffolds while staining for PECAM-1 (platelet endothelial cell adhesion molecule) displayed typical localization at cell-cell contacts. Gene expression analysis showed that endothelial cells grown on PLAGA scaffolds maintained their normal characteristic phenotype. The cell proliferation and phenotypic expression were independent of scaffold pore architecture. These results demonstrate that PLAGA sintered microsphere scaffolds can support the growth and biological functions of human endothelial cells. The insights from this study should aid future studies aimed at enhancing angiogenesis in three dimensional tissue engineered scaffolds.     

Precise centromere mapping using a combination of repeat junction markers and chromatin immunoprecipitation-polymerase chain reaction

Centromeres are difficult to map even in species where genetic resolution is excellent. Here we show that junctions between repeats provide reliable single-copy markers for recombinant inbred mapping within centromeres and pericentromeric heterochromatin. Repeat junction mapping was combined with anti-CENH3-mediated ChIP to provide a definitive map position for maize centromere 8.     

太湖梅梁湾水体中悬浮质及光谱的分布特征

在研究太湖梅梁湾水体中的悬浮质及水体中光谱分布时发现:总悬浮质中无机悬浮质含量与总悬浮质之间呈现明显的正比线形关系(R²=0.9724),而有机悬浮质与总悬浮质之间的正比线形关系较弱(R²=0.2921);在20cm处以下,总悬浮质中的藻类含量随着水深度的增加呈现下降的趋势,表面略低;有机悬浮质与叶绿素a的含量之间的线形关系较弱(R²=0.1344);光谱的分布特征是:在水体的表面,绿光最强,蓝光较强,而红光最弱;在150cm以下,红光最强,绿光较强,蓝光最弱。