单股正链RNA病毒基因组3＇非编码区功能

单股正链RNA病毒数量众多,对人、动物和植物造成很大的危害。病毒基因组3＇末端都具有一段非编码区,基因组3＇非编码区在病毒的复制过程中发挥着重要的作用。本文简单介绍了单股正链RNA病毒基因组3＇非编码区结构的预测、测定和功能的研究方法,并重点概括了不同单股正链RNA病毒3＇非编码区一级序列、茎-环结构、假结体、TLS等结构在病毒基因组的复制、转录和翻译中的调控作用以及目前存在的问题。     

Biochemical indices (RNA/DNA ratio and protein content) in studying the nutritional status of Ruditapes decussatus (Linnaeus 1758) juveniles

In this work, we investigated the nutritional status of Ruditapes decussatus juveniles under different rearing conditions, using biochemical indices (RNA/DNA ratio and protein content) and the linear instantaneous growth rate (IGR). Two experiments were conducted to study the effect of starvation, addition of substrate (sand) as rearing support and diet composition on somatic growth and biochemical indices. Results show that biochemical indices of fed juveniles were significantly (P<0.05) higher. Highest RNA/DNA values and protein content were recorded in fed juveniles reared with substrate (P<0.05); conversely, lowest values were recorded in starved ones reared without substrate. In the second experiment, the highest RNA/DNA ratio was recorded in juveniles fed with control algal mixture composed of species commonly used in bivalve nurseries [Isochrysis galbana (clone T-Iso), Chaetoceros calcitrans and Tetraselmis suecica] and lowest in those fed with algal species isolated from Tunisian coasts. This confirms the usefulness of the RNA/DNA ratio and protein content in the clam's nutritional status evaluation. This assay is useful to provide information about the nutritional status and health of the clam in field conditions.     

一种简单、有效的提取根瘤菌总RNA的方法

介绍了一种适合于根瘤菌及其它革兰氏阴性细菌的RNA提取方法。它具有简便、快速等特点,且所提取的RNA样品质量较高,可直接用于RT-PCR合成、Northern杂交等分子生物学操作。该方法具有实用性强、重复性好的特点。提取的RNA无DNA等污染物,并且其产量、纯度完全能满足分子克隆和基因表达研究的需要。该方法简单、快速、纯度高、完整性强,方便有效。     

Effect of long non-coding RNA MEG3 on invasion and migration of colorectal cancer cells

AIM: To investigate the expression of long non-coding RNA maternally expressed gene 3(MEG3) in colorectal cancer(CRC) cells, and to observe the effect of MEG3 on the invasion and migration of CRC cells. METHODS: The levels of MEG3 in human normal colon cell NCM460 and CRC cells SW48 and LoVo were detected by real- time PCR. MEG3 was over-expressed by plasmid transfection, and the effects of MEG3 on the invasion and migration of SW48 and LoVo cells were analyzed by Transwell assay and wound healing assay. The expression of matrix metalloproteinase(MMP) family proteins was determined by Western blotting. RESULTS: The level of MEG3 was down-regulated in CRC cells compared with normal colon cell NCM460. The invasion and migration of CRC cells were reduced after MEG3 over-expression. Transwell invasion and migration assays showed that the numbers of transmembrane SW48 and LoVo cells were smaller in MEG3 over-expression group than control group(CONCLUSION: The expression of MEG3 is down-regulated in CRC cells. Over-expression of MEG3 inhibits the invasion and migration of CRC cells. TIMP-2, MMP-2 and MMP-9 might play an important role in this regulation.     

Expression of Gab2 in human osteosarcoma U2-OS cells

AIM: To investigate the expression of Grb2-associated binding protein 2 (Gab2) in human osteosarcoma cells and its relationship with the invasion and metastases of human osteosarcoma cells. METHODS: The technique of small RNA interference was used to transfect human osteosarcoma U2-OS cell lines. Western blotting and RT-PCR were used to detect the protein and mRNA expression of Gab2 in transfected U2-OS cells. After transfection, through chemotaxis and invasion assays in vitro, the cell migration and invasion abilities were detected. RESULTS: After transfection, the expression of Gab2 at mRNA and protein levels in Gab2 siRNA transfected cells (SiGab2/U2-OS) was lower than that in scrambled siRNA transfected cells (Scr/U2-OS) and U2-OS cells. After stimulation with epidermal growth factor (EGF) at concentration of 10 μg/L, the migration SiGab2/U2-OS cells was significantly less than Scr/U2-OS cells and U2-OS cells (P<0.01). The number of invasion cells of SiGab2/U2-OS group was significantly lower than the other 2 control groups (P<0.01). CONCLUSION: Inhibition of Gab2 expression obviously attenuates the migration and invasion abilities of human osteosarcoma U2-OS cell line.     

Field evaluation of antisense RNA mediated inhibition of GBSS gene expression in potato

Summary Granule-bound starch synthase (GBSS) catalyses the synthesis of amylose in starch granules. Analysis of antisense RNA mediated inhibition of GBSS gene expression in large numbers of tubers from in vitro grown, greenhouse grown and field grown transgenic potato plants revealed stable and total inhibition of GBSS gene expression in one clone. In three other transgenic genotypes partial and unstable inhibition was found. In these genotypes both GBSS activity and amylose content were remarkably reduced compared with the non-transformed control genotype. No relationship was found between the level of inhibition of GBSS gene expression and yield and dry matter content.     

RNA沉默—新型的植物病毒病害防治策略

植物RNA沉默(RNAsilencing)是植物本身固有的一种抗病毒防御系统,是对病毒在复制过程中形成双链RNA(dsRNA)的特殊反应。RNA沉默介导的dsRNA干涉病毒侵染转基因抗病毒有其不足,几种体外生产dsRNA的抗病毒体系能弥补这一不足,它们与转基因表达病原RNA不同,但仍然依赖RNA沉默机制来获取病原抗性。综述了近年来发展起来的各种启动RNA沉默的dsRNA产生策略、抗病状况和存在的问题及发展前景     

ES-D3细胞nanog基因干扰载体的构建 及其干涉效果*

选择ES细胞nanog基因中4个区域合成寡聚核苷酸,构建了4个含有小鼠U6启动子的siRNA（小分子干涉RNA）表达载体并瞬时转染小鼠ES-D3细胞.半定量RT-PCR分析显示,所构建的4个siRNA表达载体中有3个可以在细胞内转录出siRNA或shRNA（短发夹RNA）,诱发RNA干涉（RNAinterference,RNAi）,能显著抑制目的基因nanog的表达;同时发现,干扰的ES-D3细胞生长48 h后,集落形态与未干扰细胞相比差别不大,但隆起不如后者明显,形态也不如后者规则,生长速度也较慢,AKP染色阳性,与未转染细胞相比没有差别.