香蕉中一个新的S-腺苷甲硫氨酸合成酶基因的克隆及采后表达分析     

贾彩红  林水玉  张建斌  刘菊华  金志强  徐碧玉 《果树学报》2009,26(3)

利用抑制消减文库从香蕉中分离到一个cDNA片断,结合RACE技术获得该基因的全长序列。该全长cDNA共含1 285个碱基,通过Blastx同源性分析结果显示它与香蕉的一个S-adenosyl-L-methionine synthase(SAMS)基因(GenBank序列号为AF004317)具有82%的碱基同源性,编码的氨基酸顺序有93%同源性,但在cDNA的5’和3’非编码区同源性较低。设计特异引物对此基因进行RT-PCR分析表明,正常成熟的香蕉果实,采后当天表达量较高,随后略有降低,至采后12 d又达到一个相对较高值后迅速下降;高锰酸钾处理的果实,整个成熟期该基因均表现一个比较高的表达量;乙烯利处理的果实,采后表达量明显下降且处在一个比较稳定的水平。  相似文献   

鲢鳙打印病豚鼠气单胞菌主要特性及系统发育学分析     

chencuizhen@.com 《勤云标准版测试》2006,(5)

对从鲢(Hypophthalmichthysmolitrix)鳙(Aristichthysnobilis)打印病病死鱼中分离到的豚鼠气单胞菌(Aeromonascaviae),进行了形态与理化特性等方面较系统的表观分类学指征鉴定。同时择代表菌株(HC960704-1)进行了16SrRNA基因的分子鉴定,测定了16SrRNA基因序列、分析了相关细菌相应序列的同源性,构建了系统发生树;HC960704-1的16SrRNA基因序列长度为1416bp(GenBank登录号:AY966888),与GenBank数据库中的气单胞菌16SrRNA基因序列的相似性在98%和100%。  相似文献   

刺槐种子中凝集素的纯化及性质研究     

王臻 《江苏林业科技》2004,31(4):1-5

从刺槐(RobiniapseudoacaciaL.)的种子中分离纯化出2种凝集素RPsAⅠ和RPsAⅡ。种子经磨碎、过滤、离心、硫酸铵分级、透析、Sepharose4B离子交换和SephadexG 150分子筛层析,即可获得在PAGE和SDS PAGE上均呈现单一蛋白染色带的凝集素纯品,分子筛层析测得RPsAⅠ和RPsAⅡ的分子量分别为132000和130000,SDS PAGE测得亚基分子量分别为38000和31000。当2种凝集素的质量浓度分别达到7 80μg/mL和15 60μg/mL时能凝集兔血红细胞,凝集活性均可被L 阿拉伯糖、D 果糖、D 半乳糖、麦芽糖抑制;热稳定性实验结果表明2种凝集素对热处理均具有一定的稳定性;当pH>9和pH<5时,它们的活性都明显降低。  相似文献   

大豆主要过敏原Gly m Bd30K蛋白单克隆抗体的制备与应用     

邹菊  刘志刚 《大豆科学》2011,30(5):723-726

利用大豆主要过敏原Gly m Bd 30K蛋白抗原表位蛋白为免疫原免疫BALB/c小鼠,取免疫小鼠脾细胞与小鼠骨髓瘤NS-1细胞融合.采用半固体培养基法和有限稀释法相结合的方法快速筛选获得稳定分泌的特异性杂交瘤细胞,用杂交瘤细胞株诱生小鼠腹水,应用蛋白A亲和层析法进行抗体纯化.采用Ig类与亚类鉴定试剂盒鉴定该单克隆抗体...  相似文献   

Impact of source tissue and ex vivo expansion on the characterization of goat mesenchymal stem cells     

Nuradilla Mohamad-Fauzi  Pablo J Ross  Elizabeth A Maga  James D Murray 《畜牧与生物技术杂志(英文版)》2015,6(1)

Background

There is considerable interest in using goats as models for genetically engineering dairy animals and also for using stem cells as therapeutics for bone and cartilage repair. Mesenchymal stem cells (MSCs) have been isolated and characterized from various species, but are poorly characterized in goats.

Results

Goat MSCs isolated from bone marrow (BM-MSCs) and adipose tissue (ASCs) have the ability to undergo osteogenic, adipogenic and chondrogenic differentiation. Cytochemical staining and gene expression analysis show that ASCs have a greater capacity for adipogenic differentiation compared to BM-MSCs and fibroblasts. Different methods of inducing adipogenesis also affect the extent and profile of adipogenic differentiation in MSCs. Goat fibroblasts were not capable of osteogenesis, hence distinguishing them from the MSCs. Goat MSCs and fibroblasts express CD90, CD105, CD73 but not CD45, and exhibit cytoplasmic localization of OCT4 protein. Goat MSCs can be stably transfected by Nucleofection, but, as evidenced by colony-forming efficiency (CFE), yield significantly different levels of progenitor cells that are robust enough to proliferate into colonies of integrants following G418 selection. BM-MSCs expanded over increasing passages in vitro maintained karyotypic stability up to 20 passages in culture, exhibited an increase in adipogenic differentiation and CFE, but showed altered morphology and amenability to genetic modification by selection.

Conclusions

Our findings provide characterization information on goat MSCs, and show that there can be significant differences between MSCs isolated from different tissues and from within the same tissue. Fibroblasts do not exhibit trilineage differentiation potential at the same capacity as MSCs, making it a more reliable method for distinguishing MSCs from fibroblasts, compared to cell surface marker expression.

Electronic supplementary material

The online version of this article (doi:10.1186/2049-1891-6-1) contains supplementary material, which is available to authorized users.  相似文献   

社会记忆视角下连州保安村民俗文化景观特色营造     

蔡健婷  赵建华  卢丹梅 《广东园林》2021,43(5):12-15

乡村民俗文化景观是社会记忆传承的重要途径,承载了乡村生活、生产、文化实践等社会记忆活动。社会记忆视角乡村民俗文化景观营造研究对增强乡村文化的地方认同,维系良好的乡土社会秩序,实现乡村特色化发展具有重要意义。以粤北地区连州市保安村为例,在调研访谈的基础上,从山水记忆、建筑记忆、民俗记忆 3 方面分析乡村民俗文化景观特质。随着社会的变迁,保安村面临山水记忆意象片段化、建筑记忆空间特色趋同化、民俗记忆传承面临危机等问题。据此,提出以保安村社会记忆载体形成“营造、传承、促进”的内生机制,构建生态记忆、生活记忆和生长记忆的社会记忆体系,营造特色化乡村民俗文化景观的实施策略。  相似文献   

Purification and characterization of stomach protease from the turbot (Scophthalmus maximus L.)     

Hai-ying Wang  Yue-jun Wang  Qing-yin Wang  Chang-hu Xue  Mi Sun 《Fish physiology and biochemistry》2006,32(2):179-188

Proteolytic activity in the different parts of the digestive tract of the turbot (Scophthalmus maximus L.) were studied in this work. One pure protease was isolated from turbot stomach and its behavior was studied. Results showed the optimum pH for proteases in the different parts of the digestive tract of the turbot were pH 2.0 for the stomach, pH 8.0 for the pylorus cecum, pH 8.0 for the foregut, pH 8.5 for the midgut, and pH 8.0 for the hindgut. The activity of proteases in the different parts of the digestive tract were in the sequence pylorus cecum protease > stomach protease > foregut protease > midgut protease > hindgut protease. The stomach protease was purified by ammonium sulfate precipitation and column chromatography on DEAE-Sepharose F.F. and Sephadex G-100. The purified enzyme gave a single band in SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Its molecular weight was found to be approximately 42,000 Da. The enzyme is stable at pH 1.0–9.0 and at temperatures below 40°C. Its activity was maximum at pH 2.0 and 40°C. When reaction time was prolonged the optimum temperature of the enzyme tended to decline. The enzyme was activated by Mn²⁺ and Cu²⁺ and inactivated by Fe³⁺. It was fully inhibited by pepstatin and partially inhibited by PMSF, TPCK, PCMB, and NBS. These results imply the enzyme is a pepsin.  相似文献   

Purification and characterization of a cysteine-like protease from the body wall of the sea cucumber <Emphasis Type="Italic">Stichopus japonicus</Emphasis>     

H. Qi  X. P. Dong  L. N. Cong  Y. Gao  L. Liu  T. Mikiro  B. W. Zhu 《Fish physiology and biochemistry》2007,33(2):181-188

The sea cucumber (Stichopus japonicus) is able to undergo autolysis in response to a variety of environmental and mechanical cues. Within the framework of a long-term study of this phenomenon we have purified a protease from the body wall of the sea cucumber by means of ion-exchange chromatography with DE-52 cellulose and gel filtration chromatography with Sephadex G-100. The final enzyme preparation was nearly homogeneous on polyacrylamide gel electrophoresis, and its molecular weight was estimated to be approximately 35.5 kDa. The purified enzyme exhibited a maximum activity for the hydrolysis of casein at pH 7.0 and 50°C and a remarkable stability at pH 4.0–7.0 and 40–60°C. Based on the inhibition and activation profiles obtained with numerous specific protease inhibitors and an activator, the protease purified from the body wall of the sea cucumber was defined to be a cysteine-like protease.  相似文献   

家畜胚胎干细胞的分离培养和鉴定     

栾黎明  李世杰  杨增明 《中国畜牧杂志》2003,39(2):48-50

家畜胚胎干细胞的分离培养和鉴定工作对于研究和利用家畜胚胎干细胞具有重大意义。本文对文献中报道的各种分离、培养和鉴定方法进行了综述 ,并介绍了几种主要家畜的胚胎干细胞在近十余年间的研究进展情况。  相似文献   

Molecular Analysis of Local Potato Cultivars from Tenerife Island Using Microsatellite Markers     

L. Barandalla  J. I. Ruiz de Galarreta  D. Rios  E. Ritter 《Euphytica》2006,152(2):283-291

We have used 19 SSR markers to fingerprint 41 local potato cultivars from 10 locations of Tenerife Island. These varieties represent relicts of the early introductions originating from South America and have been characterised previously morphologically and ecophysiologically. The SSR primers generated a varying degree of polymorphisms. A total of 67 alleles were observed, 12 of them were present in all cultivars. Several accession and group specific alleles were detected. Similarity coefficients were computed from the molecular data and cluster analyses were performed. Generally, cultivar groups with identical or related common names showed the same SSR patterns or clustered closely together. According to the molecular patterns misleading or confounded names were evident for four accessions. The dendrogram clusters were generally in good agreement with previous classifications of the accessions as Solanum tuberosum subsp. andigena, S. tuberosum subsp. tuberosum and Solanum chaucha genotypes. However in four cases the molecular patterns showed discrepancies with previous species assignments suggesting the need for a more detailed and comparative study of these accessions.  相似文献