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121.
Zhibin Sun Xingyun Qi Pinghua Li Chunxia Wu Yanxiu Zhao Hui Zhang Zenglan Wang 《Journal of Plant Biology》2008,51(1):25-34
The calcineurin B-like (CBL) proteins, comprising a large subfamily of calcium sensors in plant cells, play an important role
in many stress responses. We cloned a gene from the halophyteThellungiella halophila that is homologous toAtCBL9 inArabidopsis thaliana. The 1008-bpThCBL9 contains an ORF of 639 bp and encodes 213 amino acids, with a 5“-untranslated region of 193 bp and a 3”-untranslated region
of 176 bp. Its amino acid sequence shares high homology with AtCBLs.ThCBL9 is up-regulated by ABA, NaCI, and PEG inThellungiella leaves. Using molecular biological methods, we over-expressedThCBL9 inA. thaliana and found that this enhanced tolerances to both high salt and osmotic stress in transgenicArabidopsis.
These authors contributed equally to this work. 相似文献
122.
David Drew 《生物化学与生物物理学报:生物膜》2003,1610(1):3-10
The bacterium Escherichia coli is one of the most popular model systems to study the assembly of membrane proteins of the so-called helix-bundle class. Here, based on this system, we review and discuss what is currently known about the assembly of these membrane proteins. In addition, we will briefly review and discuss how E. coli has been used as a vehicle for the overexpression of membrane proteins. 相似文献
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Priyanka Singh Anupam Tiwari Sureshwar Prasad Singh Ravi Kumar Asthana 《Physiology and Molecular Biology of Plants》2013,19(4):521-528
Drought is the most important abiotic stress, challenging sustainable agriculture globally. For desiccation being the multigenic trait, a combination of identified genes from the appropriate organism may render crop tolerant to the water stress. Among the compatible solutes, proline plays multifaceted role in counteracting such stress. The genes encoding proline biosynthesizing enzymes, glutamate 5-kinase (G5K), and pyrroline-5-carboxylate reductase (P5CR) from the low-desiccation-tolerant cyanobacterium Anabaena sp. PCC 7120, were cloned and overexpressed in Escherichia coli BL21(DE3) individually. The recombinant E. coli cells harboring G5K, failed to exhibit enhanced desiccation tolerance relative to those with P5CR that showed increased growth/survival over the wild type. This may be ascribed to the overexpression of the reductase gene. Multiple sequence alignment showed P5CR to be conserved in all the organisms. We hypothesize that P5CR gene from high-desiccation-tolerant cyanobacteria may be adopted as the candidate for making transgenic N2-fixing cyanobacterium for paddy fields and/or crop development in future. 相似文献
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Fd:NADP+ oxidoreductase (FNR) is one of the key enzymes in photosynthetic electron transport. The gene petH encoding FNR of Synechococcus sp. PCC 7002 was cloned into the expressing vector pET-3 d' and overexpressed in E. coli. The amount of recombinant FNR (rFNR) was over 50% of the total cellular proteins. There were two forms of FNR activity, one is soluble and the other one was in the form of inclusion bodies. The soluble rFNR was purified through ion exchange chromatography and gel chromatography. The rFNR in the form of inclusion bodies was first solubilized with 6.7 mol/L urea, and then refolded into the active form in the presence of flavin adenine dinucleotide (FAD). Further purification was performed by ion exchange chromatography. The rFNR pmified from either form of the expressed product had the maximum absorption spectrum as that of the natural FNR from cyanobacteria, whose maximum absorption was at 273, 385 and 456 ran respectively. N-tenninal sequencing showed that rFNR was indeed a product of petH gene expression, rFNR could catalyze the electron transport from P700 to NADP+ in the presence of ferredoxin. The optimal pH for diaphorase activity of rFNR was 8.0 and the optimal temperature was 30 ℃. 相似文献
128.
目的:探讨人再生基因Ⅳ(REGIV)在前列腺细胞中的表达及意义.方法:构建REGIV基因的全序列过表达质粒.将全序列过表达质粒采用脂质体转染的方式转入前列腺细胞系PC-3中.应用Real-time-PCR方法检测REGIV基因mRNA表达,Westembloting检测REGIV基因蛋白质表达,MTT法分析细胞增殖活性.结果:通过荧光显微镜观察计数,细胞转染成功.REGIV基因的全序列过表达使REGIV基因mRNA的表达,蛋白表达提高,细胞增殖能力增强.结论:应用全序列过表达技术可以使前列腺癌REGIV表达水平特异性增高.前列腺癌增值能力的增强说明REGW可能与肿瘤快速增长有关. 相似文献
129.
Lovastatin (lova), a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor, can induce differentiation in cancer cells at low concentration, thus having potential to be used as an auxiliary agent in cancer therapy. However, biological networks associated with the differentiation effect of lova have not been elucidated. To investigate molecular mechanisms of lova, the present study was aimed at proteomics and bioinformatics analyses on anaplastic thyroid cancer cell line ARO differentiated with low concentration of lova. Thyroid differentiation was induced by treating ARO cells with 25 μM of lova and confirmed by checking upregulation of some thyroid differentiation markers. Gel-based proteomics analysis was then performed to identify proteins differentially expressed between undifferentiated and lova-differentiated ARO cells. Bioinformatics analysis was finally performed to estimate biological networks regulated by lova. Our results showed that lova impacted on proteins involved in protein folding, biomolecule metabolism, signal transduction, protein expression and protein degradation. Specifically, transfecting ARO cells with plasmid DNA encoding flotillin 1 (FLOT1) up-regulated the thyroid differentiation markers, indicating that FLOT1 might at least partially mediate the lova-induced thyroid differentiation. These data may shed light on the mechanism underlying lova-induced re-differentiation of thyroid cancer, and give a rationale for clinical use of lova as an auxiliary agent in cancer therapy. 相似文献
130.
The inulinase gene cloned from the marine-derived yeast Pichia guilliermondii strain 1 was expressed in Pichia pastoris X-33 and the conditions for overexpression of the inulinase were optimized. After the optimization of the conditions for production of the recombinant inulinase, 286.8 ± 5.4 U/ml and 8873 ± 55.3 U/mg of the recombinanat inulinase in the supernatant of the culture of 2-l fermentor were attained at 120 h of the fermentation and fermentation efficiency was 13.04 μg ± 0.4 of protein/ml/d. The recombinant inulinase was purified and characterized. The molecular weight of the purified recombinant inulinase was 57.6 kDa, which was higher than that of the native iunlinase. The optimal pH and temperature of the purified recombinant inulinase were 6.0 and 60 °C, respectively. Other biochemical characteristics of the purified recombinant inulinase were the same as those of the native inulinase produced by the marine-derived P. guilliermondii strain 1. The purified recombinant inulinase also had high exoinulinase activity. Therefore, the recombinant inulinase may have highly potential applications in food and pharmaceutical industies. 相似文献