企业文化基因及其识别实证研究

企业文化评价与识别研究是管理学科的热点和难点问题.在回顾企业DNA研究应用的基础上,提出并界定了企业文化基因概念,对企业文化基因识别进行了实证研究.     

一种简便有效的PCR扩增片段直接回收克隆方法

作者提供一种简便快速的克隆方法,即直接对琼脂糖凝胶电泳分离的PCR产物进行克隆,可获得较高的阳性克隆率.结果显示,300～700 bp大小的片段阳性克隆率为70.72%,300 bp以下和大于700 bp的片段阳性克隆率分别达到60.38%和45.33%.该方法适用于含有大量不同大小DNA片段的PCR产物并需要同时对其进行回收和克隆的样品,不仅可以大大简化实验过程,也可以降低假阳性出现的机会.     

Construction and anti-tumor effects of recombinant fowlpox virus expressing Newcastle disease virus hemagglutinin-neuramidinase gene

Hemagglutinin-neuramidinase (HN), a Newcastle disease virus-derived protein, not only mediates receptor recognition but also possesses neuraminidase (NA) activity, the ability to cleave a component of those receptors, N-acetylneuraminic acid (NAcneu, sialic acid). It is known that this protein in mammalian species, including human beings, has interesting anti-neoplastic as well as immune stimulating properties. To explore the use of the HN gene in cancer gene therapy, we constructed a recombi-nant fowlpox virus expressing the HN protein (vFV-HN) and compared the anti-tumor activity of the recombinant virus with that of wild-type fowlpox virus (FPV) in vivo and in vitro. Here we found that although B16 cells were somewhat resistant to the basal cytotoxic effect of wild-type fowlpox virus, infection with vFV-HN caused a pronounced cytotoxic effect and, the survival of tumor-bearing mice immunized with vFV-HN was significantly increased compared with the survival of mice immunized with the FPV alone. Furthermore, the immunization of mice with vFV-HN elicited a B16 tumor-specific cytotoxic T lymphocyte (CTL) response and clonal expansion of both CD4 and CD8 T cell populations in vivo. In addition, T cells from lymph nodes of mice vaccinated with vFV-HN secreted high levels of the Th1 cytokine IL-2 and IFN-γ, indicating that the regression of tumor cells is related to a Th1-type dominant immune response. These results demonstrate that vaccination with vFV-HN may be a potential strategy for cancer gene therapy.     

枝状枝孢霉MD2的Unigene A03231的克隆及原核表达分析

为研究真菌紫杉醇合成相关基因的功能,从产紫杉醇真菌枝状枝孢霉MD2中克隆了Unigene A03231的DNA和c DNA全长序列(918 bp).结果表明:该基因不含内含子,其编码产物含305个氨基酸,与短链脱氢酶/还原酶一致性为64%,预测蛋白分子量为33071.5 Da,不存在信号肽,含有2个跨膜结构.Unigene A03231以单拷贝形式存在于基因组,在0~30 h内其表达水平随茉莉酸甲酯诱导表现为先升高后降低.构建了该基因的原核表达菌株,初步优化了其在大肠杆菌BL21中的诱导表达条件为:诱导温度28℃,IPTG浓度0.6 mmol/L,诱导时间8 h.为深入分析该基因的催化功能奠定了基础.     

Phylogenetic position of Schnabelia, a genus endemic to China： Evidence from sequences of cpDNA matK gene and nrDNA ITS regions

The chloroplast gene matK and the internal transcribed spacers (ITS) of the nuclear ribosomal DNA from Schnabelia, a genus endemic to China, and 6 genera of Verbenaceae and 13 genera of Lamiaceae were sequenced. The phylogenetic signal and validity outgroups were measured and evaluated by means of the relatively apparent synapomorphy analysis (RASA). Independent and combined phylogenetic analyses for the matK and ITS sequences were performed using the maximum parsimony (MP), neighbor-joining (NJ) and maximum likelihood (ML) methods, indicating that Schnabelia oligophylla and Caryopteris terniflora form a sister-group relationship. The Caryopteris complex is not shown to be a monophyly because Trichostema, C. paniculata and C. forrestii are paraphyletic to the clade containing the remaining members of the complex. A monophyly of Ajugoideae proposed by Cantino et al., including 8 genera in this study, is strongly supported and the closest relatives of Schnabelia are in the Ajugoideae (Lamiaceae), especially near Caryopteris terniflora. The polygenetic analyses also showed that the genera of Lamiaceae and Verbenaceae sampled in this tudy are phylogenetically mixed and the genus Avicennia is distant to other genera of Verbenaceae. RASA and combined analysis can be used as effective approaches to determining the relationships among phylogenetically complex groups.     

Research advances in gene therapy for Parkinson's disease

During the long period of time when people have been seeking for an effective therapeutic method for PD, the gene therapy has shown greater and greater advantages over other methods. It can be performed mainly in two ways: ex vivo and in vivo. With the former, TH gene as well as some neurotrophic factor genes (such as GDNF, BNDF genes) can be invited to some cell lines or primary cells thus forming engineered cells and then implanting them into brain. While with the latter, viral vectors including HSV-1, Ad, AAV that can be utilized to construct recombinant viruses, or non-virus vectors can be used to delived DNA into brain directly. The present review summarizes the recent research advances in the gene therapy for PD, and it is reasonable for us to predict a notable progress in prevention and treatment for PD in the next decade.     

Chromosomal location of yellow rust resistance gene(s) inTriticum aestivum-Lophopyrum elongatum substitution lines

A set ofT. aestivum-L. elongatum substitution lines were studied on yellow rust resistance at seedling stage, inheritance of the resistance and esterase-5 (Est-5) analysis. The results demonstrated thatL. elongatum carried a new yellow rust resistance gene(s), which was dominant and located on chromosome 3E ofL. elongatum. The biochemical locus encoding Est-5 was also located on chromosome 3E, and was tentatively named Est-E5, which was co-segregant with theYr gene(s) in wheat background. In addition, the transmission frequencies of chromosome 3E in 3A/3E and 3D/3E hybrids selfing were significantly higher than that of chromosome 3E in 3B/3E hybrid, which was probably due to the difference on genetic relationships among A, B, D and E genomes.     

红霉素抗性基因ermE在链霉菌中的表达及抗性测定

红霉素抗性基因ermE编码一种甲基化酶,可以对50S核糖体亚基上的23SrRNA进行甲基化修饰．甲基化作用可能引起核糖体构型变化,阻碍红霉素与50S核糖体亚基的结合,从而使宿主获得抗性．研究中将来自红色糖多孢菌（Saccharopolyspora erythraea）的ermE基因克隆到链霉菌高拷贝表达栽体pUWL201和plJ6021上组成型启动子ermEp＊和硫链丝菌素诱导型启动子PfipA的下游,构建出了链霉菌表达质粒pKIM4,pKIM7和pKIM8．将pKIM4,pKIM7和pKIM8分别转入S．lividans链霉菌中,每种转化子都能表现出明显的红霉素抗性,并且ermE基因在TK64／pKIM8转化子中还实现了高效表达．     

解酯耶氏酵母产γ-癸内酯的研究进展

介绍了微生物通过β-氧化转化蓖麻油酸产生γ-癸内酯的机理和国内外对解脂耶罗维亚酵母在产γ-癸内酯方面的一系列相关研究进展,同时介绍了本研究组应用同源重组基因敲除和自克隆技术对解脂耶罗维亚酵母进行的某些基因遗传改良工作,通过本研究组的工作,应用解脂耶罗维亚酵母生产γ-癸内酯的量有了更大提高.探讨了高产γ-癸内酯酵母菌株的构建思路,以期有助于今后的研究工作.