拟南芥JR1基因片段的克隆及分析

根据拟南芥CBF基因序列的保守区设计合成一对特异引物,以菠菜基因组DNA为模板,采用PCR扩增的方法扩出一条DNA特异片段并克隆到pUC18载体中。用酶切和测序分析法对克隆片段进行鉴定并进一步进行核苷酸序列分析。序列测定该片段长为430bp。OMIGA2.0软件分析结果表明,该片段与已报道的拟南芥JR1序列的一致性为99.1%。     

新生鼠大脑皮质和中脑多巴胺神经元原代培养中NGF诱导c-jun mRNA表达的研究

用神经生长因子（nervegrowthfactor,NGF）分别处理原代培养的新生大鼠大脑皮质和中脑腹侧部神经元,应用免疫组织化学和原位杂交双重标记方法,观察不同时间NGF处理的神经元表达原癌基因c－junmRNA的情况。结果发现,神经特异性烯醇化酶（neuronspecificenolase,NSE）阳性的大脑皮层神经细胞在NGF处理15分钟即可表达c－junmRNA,2小时达高峰,4小时后开始下降,到8小时后基本消失．未经NGF处理的大鼠大脑皮层神经细胞不表达c－junmRNA;酪氨酸羟化酶（tyrosinehydroxylase,TH）阳性的中脑多巴胺能（Dopaminergic,DA）神经元经NGF处理也不表达c－jun基因。提示NGF与其受体结合可以激活神经细胞快速,短暂的一过性表达c-jun基因,作为第三信使,调节从细胞质膜到核的信号传递,同时也间接证明了新生大鼠大脑皮层神经细胞膜上存在神经生长因子受体（nervegrowthfactorreceptor,NGFR）,而中脑DA神经细胞对NGF无应答反应。     

Mapping of B-cell epitopes of the human hepatitis B virus X protein.

The immune response to the X protein of human hepatitis B virus (HBV) was studied by epitope mapping by using a set of MS2-HBx fusion proteins and synthetic peptides. Antibodies in sera of patients with acute and chronic HBV infection showed a multispecific immune response. Each serum contained antibodies to a different set of epitopes, which taken together cover most of the HBx sequence. Some of the epitopes were detectable only by immunoblotting with fusion proteins; others were detectable only by an enzyme-linked immunosorbent assay (ELISA) with synthetic peptides. The carboxy-terminal half of the HBx protein was preferentially recognized by antibodies from patients with chronic hepatitis and contained a short immunodominant antigenic region with at least two major nonoverlapping epitopes. Anti-HBx antibody titers as revealed by peptide ELISAs were highest and most frequent in patients with chronic hepatitis and usually low in acutely infected patients and asymptomatic carriers. The data demonstrate a remarkable qualitative and quantitative heterogeneity of the humoral HBx immune response which can be monitored by HBx-specific peptide ELISAs. Such tests may become useful diagnostic tools.     

卧龙自然保护区大熊猫生境恢复过程研究

保护与恢复生境是有效保护大熊猫的重要途径,通过样方调查法研究了卧龙自然保护区大熊猫（Ａｉｌｕｒｏｐｏｄａ　ｍｅｌａｎｏｌｅｕｃａ）恢复生境的群落结构特征,共调查了原始生境、２０世纪２０一３０年代砍伐后自然恢复生境、４０一５０年代砍伐后自然恢复生境、７０年代以后自然恢复生境以及６０一７０年代人工林等５个生境类型,２１个样方。研究结果表明：各生境类型的物种丰富度、物种多样性、植株数（高度＞５ｍ）、乔木层的平均胸径和最大平均胸径,以及大熊猫的生境成熟度都存在显著差异,竹子的生物量及更新能力也有一定差异。人工林与原始生境的群落相似性低于其他自然恢复生境与原始生境的群落相似性。研究发现,大熊猫生境恢复包括大熊猫可食竹类资源的恢复以及生境群落结构的恢复。可食竹类资源恢复所需时间相对较短,仅需约２０一３０ａ;生境的植物群落结构的恢复则要长的多,一般恢复时间５０ａ左右才能成为大熊猫的适宜生境,恢复时间为７０一８０ａ的生境与原始生境的群落结构已十分接近。通过人工造林恢复生境,无论从竹子资源的恢复,还是从植物群落结构的恢复方面,均不是一种有效地恢复大熊猫生境的方式。     

利用絮凝进行微藻采收的研究进展

微藻可生产不饱和脂肪酸及色素等多种高附加值产品,同时也可用来生产可再生清洁能源如生物柴油等,具有良好的应用前景。但是,目前微藻细胞的采收成本高居不下,已成为限制微藻生物技术大规模应用的重要因素之一。与其他方法相比,絮凝采收成本低、操作简便,是很有应用前景的采收方法。本文综述了国内外利用化学絮凝、物理絮凝及生物絮凝等方法对不同微藻细胞进行采收的研究,重点对生物絮凝方法进行了总结。利用微生物絮凝剂及微藻细胞的自絮凝进行微藻生物量的回收,是微藻采收技术中环境友好、低成本和行之有效的新方法之一。     

Generation of ΔF508-CFTR T84 cell lines by CRISPR/Cas9-mediated genome editing

Objectives: To provide a simple method to make a stable ΔF508-CFTR-expressing T84 cell line that can be used as an efficient screening model system for ΔF508-CFTR rescue. Results: CFTR knockout cell lines were generated by Cas9 with a single-guide RNA (sgRNA) targeting exon 1 of the CFTR genome, which produced indels that abolished CFTR protein expressions. Next, stable ΔF508-CFTR expression was achieved by genome integration of ΔF508-CFTR via the lentivirus infection system. Finally, we showed functional rescue of ΔF508-CFTR not only by growing the cells at a low temperature, but also incubating with VX-809, a ΔF508-CFTR corrector, in the established T84 cells expressing ΔF508-CFTR. Conclusions: This cell system provides an appropriate screening platform for rescue of ΔF508-CFTR, especially related to protein folding, escaped from endoplasmic-reticulum-associated protein degradation, and membrane transport.     

APC/C-Cdh1: From cell cycle to cellular differentiation and genomic integrity

Anaphase-promoting complex/cyclosome (APC/C) is a multifunctional ubiquitin-protein ligase that targets various substrates for proteolysis inside and outside of the cell cycle. The activation of APC/C is dependent on two WD-40 domain proteins, Cdc20 and Cdh1. While APC/Cdc20 principally regulates mitotic progression, APC/Cdh1 shows a broad spectrum of substrates in and beyond cell cycle. In the past several years, numerous biochemical and mouse genetic studies have greatly attracted our attention to the emerging role of APC/Cdh1 in genomic integrity, cellular differentiation and human diseases. This review will aim to summarize the recently expanded understanding of APC/Cdh1 in regulating biological function and how its dysfunction may lead to diseases.Key words: APC/C, Cdh1, proteolysis, genomic integrity, signal transduction, differentiation, tumorigenesis