Novel promoter/transactivator configurations for macrolide- and streptogramin-responsive transgene expression in mammalian cells

Background

The recently developed heterologous macrolide‐ (E.REX system) and streptogramin‐ (PIP system) responsive gene regulation systems show significant differences in their regulation performance in diverse cell lines.

Methods

In order to provide optimal regulation modalities for a wide variety of mammalian cell lines, we have performed a detailed analysis of E.REX and PIP systems modified in (i) the transactivation domains of the antibiotic‐dependent transactivators, (ii) the type of minimal promoter used, and (iii) the spacing between the operator module and the minimal promoter.

Results

These novel E.REX and PIP regulation components showed not only dramatically improved regulation performance in some cell types, but also enabled their use in cell lines which had previously been inaccessible to regulated transgene expression.

Conclusions

Due to their modular set‐up the novel E.REX and PIP regulation systems presented here are most versatile and ready for future upgrades using different cell‐specific key regulation components. Copyright © 2002 John Wiley & Sons, Ltd.

     

贵州鼠尾草化学成分的研究

从贵州鼠尾草95%乙醇提取物分离得到8个化合物,通过理化性质及波谱方法分别鉴定为正三十三烷(1),十二烷酸十四烷酯(2),β-谷甾醇(3),胡萝卜苷(4),乌苏酸(5),2 a,3β-二羟基乌苏酸(6),白桦酸(7)和迷迭香酸(8)。以上化合物均首次从该植物中分离得到。     

Identification of Pip4k2β as a mechanical stimulus responsive gene and its expression during musculoskeletal tissue healing

To investigate the mechano-transduction system of cells, we identified genes responsive to a cyclic mechanical stimulus. MC3T3.E1 cells were cultured on a computer-controlled vacuum-pump-operated device designed to provide a cyclic mechanical stimulus. A maximum elongation of 15% of membrane at 10 cycles/min (3 s extension followed by 3 s relax per cycle) was repeated for 48 h. By means of a differential display, the gene expression pattern of cells exposed to the stimulus was compared with that of unexposed cells. As a result, a gene fragment that was exclusively expressed in mechanically stressed cells was identified. By using expressed sequence tag walking together with the oligo-capping method, this gene was identified as phosphatidylinositol 4-phosphate 5-kinase type II β (initially known as Pip5k2β but now reclassified as Pip4k2β). The specific up-regulation of Pip4k2β upon mechanical stimulus was also confirmed by using another apparatus, viz. a computer-controlled linearized-stepping motor system. To examine the involvement of the cyclic mechanical stimulus in the regulation of Pip4k2β expression in musculoskeletal tissue, we created an Achilles tendon transection model in rabbits. The temporal expression of Pip4k2β was assessed by means of a quantitative reverse-transcribed polymerase chain reaction. In the gastrocnemius muscle, expression of Pip4k2β rapidly decreased 1 week after transection but was restored to normal levels at 4 weeks. In the Achilles tendon, however, expression remained decreased until 4 weeks after transection. We suggest that the expression of Pip4k2β can be used as a marker for cells receiving a suitable mechanical stimulus. This study was supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science, Sports and Culture of Japan.     

中国大戟科一新归化种——硬毛巴豆

首次报道了大戟科(Euphorbiaceae)巴豆属硬毛巴豆(Croton hirtus L'Hér.)在中国的归化新记录,并提供该种的特征描述和形态图。凭证标本保存于中国科学院华南植物园标本馆(IBSC)。     

麦红吸浆虫滞育习性研究

1986～1991年研究了麦红吸浆虫的滞育习性,提出了该由主动延长滞育和被动延长滞育的两种形式,探讨了土壤湿度、温度对延长滞育的影响,从体重、羽化率、卵胚数、死亡率等方面测定了延长滞育幼虫的生殖势能,文章最后讨论了麦红吸浆虫多峰羽化的特点和延长滞育的生物学意义。     

桉树新害虫——曲线纷夜蛾的生物学和生态特性研究

曲线纷夜蛾Polydesma boarmoides Guenée是我国桉树食叶害虫,已在部分桉树种植区造成严重危害。国内外仅在区域昆虫种类调查中发现并提及该虫,而未见其生物学特性、生态学特性和发生规律等方面的相关研究报道,因此,有必要对该虫进行更系统、深入的研究,为防控该虫提供依据。笔者2013年5月首次发现该虫在广东省紫金县危害桉树林,2016年5月再次发现该虫在茂名市危害桉树林。调查、研究结果表明:(1)曲线纷夜蛾起始取食灌木叶片直至叶片被取食殆尽后,逐步转移至桉树林继续取食桉树叶片;(2)曲线纷夜蛾在广东地区1年发生4代;(3)曲线纷夜蛾3-5龄幼虫在桉树林间呈现聚集分布,且聚集密度呈现随种群密度增大而增大的规律,而在桉树树冠上则呈均匀分布。     

Q-band electron paramagnetic resonance study of nitrosylprotoheme dimethyl ester complexes with N-,O-, and S-donor ligand as model systems for nitrosylhemoproteins

Electron paramagnetic resonance (epr) spectra (at X- and Q band frequency) of nitrosyl(proto-porphyrin IX dimethyl ester) iron( II) complexes with a trans axial ligand of nitrogen-, oxygen-, and sulfur-donor ligand, in the trozen glass state at 77°K, have been investigated in order to understand the epr spectra of nitrosylhemoproteins. The Q-band spectra resolved the spectral features more clearly than the X-band spectra and distinctly exhibited two groups of absorptions, which were attributable to two molecular species. Significant relations were found between two g values (e.g., g_x-g_z, g_x-g_y) and between the g value and the degree of the hyperfine splitting in central absorption. The epr parameters were not very sensitive to the π-bonding ability of the axial ligand, but registered the steric interaction of the axial ligand with porphynnato core. These findings can be utilized in the characterization of an axial ligand trans to the nitrosyl group in nitrosylhemoproteins.     

艾叶多糖提取率影响因素分析及提取条件优化

艾叶是菊科(Compositae)植物艾(Artemisia argyi Lévl.et Vant.)的干燥叶,为常用中药材之一,药用历史悠久,具有苦燥辛散、理气血、温经脉、逐寒湿、止冷痛等功效,为重要的妇科用药,可用于治疗脘腹冷痛、经寒不调、宫冷不孕等证.此外,利用艾叶预防瘟疫已有悠久的历史.现代药理研究表明:艾叶是一种广谱抗菌抗病毒中药,对多种病毒和细菌有抑制和杀伤作用,对呼吸系统疾病以及心血管等慢性疾病有一定的防治作用[1-3].因此,将艾叶中的有效成分分别提取和纯化并加以分析以及功能检测,具有深远意义.