九株根霉可溶性蛋白和三种酶的电泳图谱比较研究

本文报导了根霉属(Rhizopus) 9个菌株天然态及解聚态可溶性蛋白、酯酶同工酶、葡萄糖淀粉酶和SOD电泳图谱的比较研究。结果表明：可溶性蛋白图谱和酯酶同工酶谱能显示五种已知供试菌种间的差异,尤其酯酶同工酶谱还能显示米根霉两个供试菌株之间的微小差异。经综合分析全部试验结果后得出的系统树图显示了9个供试菌株间的亲缘关系,并为未知菌株F1(BR12)和Q303提供了鉴定和命名依据。文中首次报导了根霉的SOD同工酶,并对蛋白质和酶电泳图谱用于根霉分类研究进行了讨论。     

精子作载体的转基因鱼研究

本文报道了以精子为载体将美洲拟蝶抗冻蛋白基因导入罗非鱼卵,构建转基因鱼的方法,此法简单易行。斑点杂文和ＳｏｕｔｈｅｒｎＢｌｏｔ杂交结果表明,外源基因的整合率为１８．１％,与其它方法构建转基因鱼的外源基因整合率相近。     

降血脂药物（DEHP）对酰基CoA氧化酶基因反式作用因子的影响

从对照和用ＤＥＨＰ处理的大鼠肝脏提取核蛋白,以含酰基ＣｏＡ氧化酶（ＡＯＸ）基因表达调控部位的ＤＮＡ片段和该基因的不同蛋白结合位点的ＤＮＡ片段作为核蛋白结合反应的探针,通过凝胶电泳迁移率改变实验和Ｓｏｕｔｈｗｅｓｔｅｒｎ印迹分析检查了ＤＥＨＰ对ＡＯＸ基因反式作用因子的影响。结果表明,降血脂药物ＤＥＨＰ可显著增加ＡＯＸ基因反式作用因子的含量和（或）与基因的结合活性,在转录水平上促进基因的表达。     

A phenotypic host range alteration determines RD114 virus restriction in feline embryonic cells.

We have characterized the restriction mechanism for RD114 virus replication in embryonic feline cells (FeF). By comparing growth properties of the virus in FeF cells with its behavior in a fetal feline glial cell line (G355) permissive for RD114, we showed that both cell lines were readily infectible by virus grown in permissive cells and that no significant differences in viral integration or viral RNA expression could be detected. However, analysis of viral protein expression revealed differences in viral env gene processing in the two cell types. Envelope precursor pR85 was produced, but the expected processed gp70 product was detectable only in permissive (G355) cells. An envelope product of 85 kDa was packaged into virions produced by FeF cells, while virions produced by G355 cells contained the expected RD114 gp70. While the gp85 env-containing virions were infectious for permissive G355 cells, they were unable to infect FeF cells. The block to infection by the gp85-containing particles in FeF cells could be abrogated by treatment with the glycosylation inhibitor tunicamycin. Our results indicate that restriction of RD114 virus involves a novel mechanism dependent on two factors: altered glycosylation of the envelope to a gp85 form and an altered RD114 receptor in FeF cells.     

湖羊瘤胃微生物GH9家族葡聚糖酶基因IDSGLUC9-25的表达与功能表征

【目的】葡聚糖酶是饲用添加剂的重要成分,本研究旨在从湖羊消化道微生物中挖掘性质优良的GH9家族葡聚糖酶基因,用于研发新型饲用酶制剂。【方法】从湖羊瘤胃微生物cDNA中扩增IDSGLUC9-25基因,在大肠杆菌中进行异源表达,对重组蛋白进行诱导表达和纯化,研究重组蛋白的酶学性质和底物水解模式。【结果】IDSGLUC9-25基因编码527个氨基酸,包含一个CelD_N结构和一个GH9家族催化结构域;重组蛋白rIDSGLUC9-25分子量约为62.7 kDa,最适反应温度和pH分别为40℃和6.0,在30-50℃下活性较高,在pH 4.0-8.0范围内能够保持较高的稳定性,经pH 4.0-8.0缓冲液处理1 h后残余活性均大于90%;底物谱分析表明,rIDSGLUC9-25能催化大麦β-葡聚糖、苔藓地衣多糖、魔芋胶和木葡聚糖,比活性分别为(443.55±24.48)、(65.56±5.98)、(122.37±2.85)和(159.16±7.73) U/mg;利用薄层色谱法(thin layer chromatography, TLC)和高效液相色谱法(high performance liquid chromatography, HPLC)分析水解产物发现,rIDSGLUC9-25降解大麦葡聚糖主要生成纤维三糖(占总还原糖64.19%±1.19%)和纤维四糖(占总还原糖26.24%±0.12%),催化地衣多糖主要生成纤维三糖(占总还原糖78.46%±0.89%)。【结论】本研究报道了一种来自密螺旋体属细菌的内切β-1,4-葡聚糖酶IDSGLUC9-25 (EC 3.2.1.4),能高效催化多糖底物生成纤维三糖和纤维四糖,为研发饲用酶制剂和制备低聚寡糖建立基础。     

骨缘当归的化学成分

骨缘当归Angellica cartilaginomarginata var.foliata Yuan et Shen,别名:山藁本、野芹菜（江苏）,仅分布于江苏和浙江。在江苏镇江等地用干燥全草作山藁本入药,其化学成分未见报道。 样品采自江苏省句容县。将干根磨粉,提取物经中性氧化铝层析,根据色谱及光谱鉴定为两种脂肪酸、四种已知香豆素和一种植物甾醇即:正葵酸（n-capric acid）、月桂酸（lauric acid）、骨缘当归素（cartilaginomarginadin）、蝉翼素（pteryxin）、白花前胡素F（praeruptorin F）、佛手柑内酯（bergapten）及β-谷甾醇（β-sitosterol）。     

UreB immunodominant epitope-specific CD8+ T-cell responses were beneficial in reducing gastric symptoms in Helicobacter pylori-infected individuals

Background and aims

Although Helicobacter pylori is recognized as an extracellular infection bacterium, it can lead to an increase in the number of CD8⁺ T cells after infection. At present, the characteristics of H. pylori antigen-specific CD8⁺ T cells and the epitope response have not been elucidated. This study was focused on putative protective antigen UreB to detect specific CD8⁺ T-cell responses in vitro and screen for predominant response epitopes.

Methods

The PBMCs collected from H. pylori-infected individuals were stimulated by UreB peptide pools in vitro to identify the immunodominant CD8⁺ T-cell epitopes. Furthermore, their HLA restriction characteristics were detected accordingly by NGS. Finally, the relationship between immunodominant responses and appearance of gastric symptoms after H. pylori infection was conducted.

Results

UreB-specific CD8⁺ T-cell responses were detected in H. pylori-infected individuals. Three of UreB dominant epitopes (A-2 (UreB_443–451: GVKPNMIIK), B-4 (UreB_420–428: SEYVGSVEV), and C-1 (UreB_5–13: SRKEYVSMY)) were firstly identified and mainly presented by HLA-A*1101, HLA-B*4001 and HLA-C*0702 alleles, respectively. C-1 responses were mostly occurred in H. pylori-infected subjects without gastric symptoms and may alleviate the degree of gastric inflammation.

Conclusions

The UreB dominant epitope-specific CD8⁺ T-cell response was closely related to the gastric symptoms after H. pylori infection, and the C-1 (UreB_5-13) dominant peptides may be protective epitopes.