摘要：
Expression profiles of 12 class A rice heat shock transcription factor genes (OsHsfAs) were analyzed by semi-quantitative reverse transcriptase polymerase chain reaction. The OsHsfA genes exhibited tissue-specific expressions under normal condition. OsHsfA1a, A2d, and A9 were predominantly expressed in young spike. Expression responses of the 12 OsHsfAs under abiotic stresses were analyzed in the shoots of rice seedling. Most OsHsfA genes responded quickly to heat stress except for OsHsfA1a, A3, and A9 which were almost unaffected. In particular, OsHsfA2a expression in response to heat stress was highest among the heat shock factors examined. However, the majority of the increased OsHsfAs expression responses to salt, polyethylene glycol (PEG), and cold treatments primarily occurred during the later stages (3 to 24 h) of stress exposure. Furthermore, most of OsHsfA gene expressions were little affected and only a few (OsHsfA3, A4d, A7, and A9) genes had slow responses to cold treatment. The results indicate that the transcript levels of OsHsfAs during heat stress exposure were distinct from those of plants subjected to salt, PEG, and cold stresses, suggesting that there might be different regulatory networks between heat and non-heat stress.
摘要：
The y + LAT1 gene is a member of solute carrier family that encodes a protein, which plays a significant function in transport of neutral and dibasic amino acids. The Tibetan pig is an excellent local variety in China, which is able to tolerate and survive under the adverse conditions. The study cloned the cDNA sequence of Tibetan pig y + LAT1 (GenBank accession #EU390782), and evaluated the tissue distribution and developmental expression of y + LAT1 mRNA and protein in different parts of intestine in Tibetan suckling piglets. The results show that the y + LAT1 cDNA encodes 511 deduced amino acid residues and 12 putative transmembrane domains, which can be widely detected in many organs in Tibetan piglets. Expression patterns were similar in jejunum and ileum, where the mRNA and protein level was decreased before the suckling period and increased until Day 35, duodenum changed reversely. The jejunum posterior is the predominant expressional tissue in all days. These results indicated that the ontogenesis expression of y + LAT1 was not only differentially regulated by early development age but also differentially with distributed tissues along the small intestine.
作者机构：
[曹亮; 徐菲; 魏宝阳; 佟志远] Key Laboratory of Modernization of Chinese Materia Medica, College of Pharmacy, Hunan University of Traditional Chinese Medicine, Changsha 410208, China;[黄丹] Hunan Academy of Traditional Chinese Medicine, Changsha 410013, China;[李顺祥] Key Laboratory of Modernization of Chinese Materia Medica, College of Pharmacy, Hunan University of Traditional Chinese Medicine, Changsha 410208, China, Hunan Academy of Traditional Chinese Medicine, Changsha 410013, China
通讯机构：
[Cao, L.] K;Key Laboratory of Modernization of Chinese Materia Medica, College of Pharmacy, Hunan University of Traditional Chinese Medicine, China
摘要：
In this study, we used native gradient-polyacrylamide gel electrophoresis and electroelution (NGGEE) to purify enzymatic proteins from Trichoderma koningii AS3.2774. With this method, we purified eight enzymatic proteins and classified them to the cellulase system by comparing secretions of T. koningii in inductive medium and in repressive medium. It resulted in 24-fold beta-glucosidase (BG) purification with a recovery rate of 5.5%, and a specific activity of 994.6 IU mg(-1) protein. The final yield of BG reached 8 mu g under purifying procedure of NGGEE We also identified BG using the enzyme assay with thin-layer chromatography and MALDI-TOFMS. This BG had one subunit with a molecular mass of 69.1 kDa as determined by sodium dodecylsulfate-polyacrylamide gel electrophoresis. The hydrolytic activity of the BG had an optimal pH of 5.0, an optimal temperature of 50 degrees C. an isoelectric point of 5.68 and a K-m for p-nitrophenyl-beta-D-glucopyranoside of 2.67 mM. Taken together, we show that NGGEE is a reliable method through which mu g grade of active proteins can be purified. (C) 2010 Elsevier B.V. All rights reserved.
摘要：
The present study describes a rapid and sensitive HPLC method for the quantification of huperzine A (HupA) in Huperzia crispata (Huperziaceae). The sample extraction and preparation involved a simple, time-saving, single-solvent extraction, with each sample being analyzed within 12 min. The mobile phase was ammonium acetate (0.1 M, pH 6.0)-methanol (64 + 36, v/v) at a flow rate of 1.0 mL/min. Detection was at 308 nm. The calibration curve was linear from 0.049 to 7.84 μg (R<sup>2</sup> = 0.9997), with intraday and interday precision RSD of less than 2%. The extraction recovery rate was over 98.49%. Quantification of HupA was performed using this modified method, and the content of HupA was 1.86 times higher in the whole plant of H. crispata (218.17 ±1.55 μg/g) than in that of H. serrata (117.03 ±2.97 μg/g). In the whole plant of H. crispata, HupA mainly accumulated in the actively growing shoot tips, the apical bud, and the 10 youngest leaves, reaching 455.23 ±2.97 μg/g. The content of HupA in the samples from sunshine-sheltered sites was 3.45 times higher than in that from sunshine-abundant sites. The satisfactory results indicate that this modified method can be applied in the quality control of large-scale Huperziaceae plant extracts and that changes should be made in the cultivation of H. crispata so as to maximize the production of HupA.
