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排序方式: 共有2235条查询结果,搜索用时 15 毫秒
31.
棉花RNA的快速提取方法 总被引:35,自引:7,他引:35
目前已发展的从植物中提取RNA的方法有多种.但由于棉花中次生物质含量较高,因此有些方法并不适合于棉花RNA的提取.本文介绍经过反复实验后改进的棉花RNA的提取方法,它具有快速、简单等特点. 相似文献
32.
采用RT-PCR方法克隆了棉铃虫蜕皮调节转录因子HaHR3基因,该基因含有1671 bp的完整开放阅读框。序列分析表明,HaHR3与多种昆虫蜕皮调节转录因子高度同源。利用生物信息学方法对HaHR3的结构特征进行分析发现,HaHR3具有蜕皮调节转录因子超家族的典型特征,包括两个锌指结构和一个DNA结合结构域,不存在信号肽序列和N糖基化位点。选择HaHR3基因的部分片段构建了RNAi中间载体pRNAi1017-HaHR3sa,再将HaHR3正反向序列亚克隆至植物表达载体pCAMBIA2300-35S-OCS,成功构建了由35s启动子调控的HaHR3基因的正反向RNA干扰载体pCAM-RNAi-HaHR3。这一载体的成功构建为下一步通过植物介导的RNA干扰技术防治棉铃虫打下了坚实的基础。 相似文献
33.
Photoperiod is commonly controlled in the commercial production of ornamental crops to induce or prevent flowering. Flower induction in short-day (SD) plants can be prevented or delayed when the natural daylength is short by providing low-intensity lighting during the dark period. A stationary high-pressure sodium (HPS) lamp with an oscillating aluminum parabolic reflector (cyclic HPS) has been developed to provide intermittent lighting to greenhouse crops. We determined the efficacy of a cyclic HPS lamp at preventing flowering in SD plants garden chrysanthemum [Chrysanthemum × grandiflorum (Ramat.) Kitam.] ‘Bianca’, pot chrysanthemum ‘Auburn’, and velvet sage (Salvia leucantha L.) relative to traditional night interruption (NI) lighting strategies. Plants were grown in a glass-glazed greenhouse at a mean daily temperature of 19.5–20.7 °C with natural SD photoperiods. NI lighting was delivered during the middle of the night (2230–0230 h) from a 600 W cyclic HPS lamp mounted at one gable end of the greenhouse or from incandescent (INC) lamps that were illuminated for the entire 4 h (CONT INC) or for 6 min every 30 min for 4 h. Plants under cyclic HPS were grown at lateral distances of 1, 4, 7, 10, or 13 m from under the lamp. Control plants were grown under an uninterrupted 15 h skotoperiod. As the distance from the cyclic HPS lamp increased from 1 to 13 m, the maximum irradiance measured during the NI decreased from 25.4 to 0.3 μmol m−2 s−1 and time to visible inflorescence (VI) and the number of nodes at VI decreased. All species had a VI within 54 d, but ≤10% of plants flowered when grown at a lateral distance of 1 or 4 m from the cyclic HPS lamp or under CONT INC. Plants grown without NI had a VI 2 to 15 d earlier and flowered 7 to 24 d earlier than plants grown at 10 or 13 m from the cyclic HPS. All garden chrysanthemums flowered under cyclic INC, whereas velvet sage and pot chrysanthemum had 15% and 35% flowering, respectively. These results indicate that a cyclic HPS lamp can be used effectively to delay flower induction and prevent flowering in these species when NI is delivered at ≥2.4 μmol m−2 s−1. 相似文献
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35.
利用DAD1反义片段转化创建菜薹可调控雄性不育材料 总被引:1,自引:0,他引:1
菜薹因为没有好的雄性不育材料或自交不亲和系,至今尚无一代杂种用于生产,根据拟南芥及白菜型油菜的花药不开裂基因DAD1的保守序列设计引物,扩增菜薹的DAD1基因片段(DAD1F),构建反义DAD1F植物表达载体,用农杆菌介导法转化菜薹,对转基因植株进行分子检测,鉴定其雄性不育性并进行育性恢复试验.克隆得到的菜薹的DAD1基因片段大小为678 bp,命名为BrcpDAD1F,其序列与拟南芥和白菜型油菜的DAD1高度同源,同源率分别为88%和99%;共得到了12株转基因植株,有6株在转录水平上得到表达,表现为雄性不育,花器官畸形,花粉活力低,萌发率不到10%,且开花后不能结角果或结空角果,或者得到极少种子但种子不萌发;用对照的花粉给转基因植株授粉可使其正常结实.以500 μmol·L-1茉莉酸甲酯处理可使其雄性不育得到恢复,花粉可以在柱头和培养基上萌发,具有受精能力。T1代可育株与不育株的比例都呈1:3分离, T2代不同株系的育性分离比例不同,有些株系继续呈1:3的分离,有些株系全是可育株或全是不育株,说明反义抑制呈单基因稳定遗传。 相似文献
36.
锦鲤保护性组织RNA提取及引物扩增研究 总被引:1,自引:0,他引:1
从养殖锦鲤各种组织中提取完整的RNA, 是分子生物学研究锦鲤应激表达、体色变异、生长发育等功能基因表达的关键实验。在不伤害养殖锦鲤个体健康的情况下, 取用鱼体的鳍条、鳞片、鳃丝、血液4种保护性组织,可以最大程度地减少对珍贵锦鲤个体的损伤, 并达到研究相关基因表达的目的。本研究利用TR izo l法提取RNA, 获得的各样品RNA条带完整、纯度高。结果表明, 4 种保护性组织提取的RNA 经过逆转录反应, 得到了高质量的CDNA。内标基因( B- actin)、热激蛋白基因( hsp70)、金属硫蛋白基因(MT)扩增得到清晰的条带, 通过NCB I数据上的BLAST比对证明, 各序列的同源性在95%以上。利用本研究的方法, 可以充分保护养殖锦鲤个体的完好性, 用于进一步的分子生物学研究。 相似文献
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38.
AIM To investigate the effect of sinomenine (SN) on the damage of human neuroblastoma SK-N-SH cells induced by 1-methyl-4-4 phenylpyridine (MPP+) and its mechanism for exploring the pathogenesis of Parkinson disease. METHODS SN was used to treat MPP+-induced SK-N-SH cells. The levels of malondialdehyde (MDA) and glutathione (GSH) in cell culture supernatants were measured by ELISA. The apoptosis was analyzed by flow cytometry. The protein expression levels of Bcl-2 and Bax were determined by Western blot. The expression levels of long noncoding RNA ANRIL and microRNA-626 (miR-626) were detected by RT-qPCR. Dual-luciferase reporter assay was used to evaluate the relationship between ANRIL and miR-626. After ANRIL small interfering RNA was transfected into SK-N-SH cells, the effects of ANRIL expression knock-down on MPP+-induced SK-N-SH cell apoptosis, the protein expression levels of Bcl-2 and Bax, and the levels of MDA and GSH in cell culture supernatants were examined. RESULTS After treatment with MPP+, the apoptotic rate, Bax protein level and ANRIL expression in SK-N-SH cells were increased (P <0.05), and the Bcl-2 protein level and miR-626 expression were decreased (P <0.05). The level of MDA in cell culture supernatants was increased (P <0.05), and the level of GSH was decreased (P <0.05). After SN treatment or ANRIL expression knock-down, decreased apoptotic rate, Bax protein level and ANRIL expression (P <0.05), and increased Bcl-2 protein level and miR-626 expression in MPP+-induced SK-N-SH cells were observed (P <0.05). The level of MDA in the cell culture supernatants was decreased (P <0.05), and the level of GSH was increased (P <0.05). CONCLUSION SN attenuates MPP+-induced damage in SK-N-SH cells by regulating ANRIL/miR-626 signaling pathway. 相似文献
39.
GUAN Cang-hai ZHAO Yu-qiao Guo Liang CHEN Yu-Zhu WANG Wei-Na JIANG Xing-ming 《园艺学报》2000,36(10):1908-1912
MNX1 antisense RNA 1 (MNX1-AS1) is a newly discovered long non-coding RNA (lncRNA), which is highly dysregulated in various carcinomas and its expression level is closely related to the overall survival and prognosis of patients. MNX1-AS1 regulates the occurrence and development of carcinomas by endogenous competitive adsorption of miRNA, regulating cell cycle, inducing epithelial mesenchymal transformation and activating multiple signaling pathways. The in-depth study of the carcinogenesis of MNX1-AS1 is useful for the early diagnosis, targeted therapy and prognostic assessment of relevant carcinomas. This article reviews the roles of MNX1-AS1 in malignant tumor. 相似文献
40.
AIM To investigate the mechanism of long noncoding RNA (lncRNA) FEZF1-AS1 regulating microRNA-363-3p (miR-363-3p) on the viability and apoptosis of lipopolysaocharide (LPS)-induced vascular endothelial cells. METHODS Human umbilical vein endothelial cells (HUVECs) were cultured in vitro . pcDNA-NC, pcDNA-FEZF1-AS1, anti-miR-NC, anti-miR-363-3p, miR-NC and miR-363-3p mimics were transfected into the HUVECs and LPS stimulation was applied for 24 h. RT-qPCR was used to detect the expression of FEZF1-AS1 and miR-363-3p. The cell viability was measured by MTT assay. The apoptotic rate was analyzed by flow cytometry. The dual-luciferase reporter experiment was used to verify the targeted regulation of FEZF1-AS1 and miR-363-3p. Western blot was used to determined the expression of cyclin D1, Ki67 and cleaved caspase-3. RESULTS Compared with control group, the expression level of FEZF1-AS1 in LPS group was significantly reduced (P <0.05), and the expression level of miR-363-3p was significantly increased (P <0.05). Compared with pcDNA-NC+LPS group, the cell viability in pcDNA-FEZF1-AS1+LPS group was significantly increased (P <0.05), the apoptotic rate was significantly reduced (P <0.05), the protein levels of cyclin D1 and Ki67 were significantly increased (P <0.05), and the protein level of cleaved caspase-3 was significantly reduced (P <0.05). Compared with anti-miR-NC+LPS group, the cell viability in anti-miR-363-3p+LPS group was significantly increased (P <0.05), the apoptotic rate was significantly reduced (P <0.05), the protein levels of cyclin D1 and Ki67 were significantly increased (P <0.05), and the protein level of cleaved caspase-3 was significantly reduced (P <0.05). Dual-luciferase reporter experiment confirmed that FEZF1-AS1 targeted miR-363-3p. Compared with miR-NC+pcDNA-FEZF1-AS1+LPS group, the cell viability in miR-363-3p+pcDNA-FEZF1-AS1+LPS group was significantly reduced (P <0.05), the apoptotic rate was significantly increased (P <0.05), the protein levels of cyclin D1 and Ki67 were significantly reduced (P <0.05), and the protein level of cleaved caspase-3 was significantly increased (P <0.05). CONCLUSION Over-expression of FEZF1-AS1 promotes the viability and inhibits apoptosis of LPS induced vascular endothelial cells by inhibiting the expression of miR-363-3p. 相似文献