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1.
曾倩  王毓舒  孙鲲  俞勇  陈波 《极地研究》2011,23(2):108-114
从北极海冰中分离筛选出一株产β-半乳糖苷酶的菌株BSi20414,经初步鉴定,该菌株归属于海单胞属(Marinomonas),命名为Marinomonas sp.BS120414.对该菌株的产酶条件实验表明,最适产酶条件为:30℃,180 rpm摇瓶培养96 h,培养基中加1.5%(w/v)乳糖,初始pH 7.0,接种...  相似文献   

2.
自北冰洋楚科奇海、加拿大海盆、格陵兰海和中国南极中山站近岸的海水、海冰和沉积物样品中分离的322株低温细菌中筛选出73株低温β-半乳糖苷酶产生菌,并对其中1株BCw006进行了研究。通过16S rDNA序列同源性分析表明,该菌株属于交替单胞菌属,定名为Alteromonas sp.BCw006。对该菌株研究表明:该菌最适生长和产酶温度为24℃;在pH6.0—8.0、含0%—6.O% NaCl的培养基条件下菌株能够较好地生长与产酶,其最适生长和产酶NaCl浓度分别为3%和1%,乳糖对β-半乳糖苷酶合成具有明显诱导作用,酵母粉和蛋白胨是菌株生长和产酶较适宜的氮源,酶的最适作用温度为35℃,0℃时能保持35℃下酶活的25%左右,表明其具有较好的适应低温特性。酶对热敏感,60℃时作用30min后酶活力下降了41%。  相似文献   

3.
采用多种培养基对北极高纬地区(土壤及海洋沉积物)真菌进行分离纯化,共得真菌65株。通过分析菌株的18S rDNA序列研究了它们的系统发育多样性,18S rDNA序列分析表明,65株真菌分属25个属,其中数量上位居第一的是青霉属(Penicillium),有菌株20株,其次为丛赤壳属(Nectria)和节枝孢属(Articulospora),各有菌株6株。同时对这65株真菌开展转化龙胆苦苷阳性菌株的筛选。以β-葡萄糖苷酶作为筛选转化菌株的标志酶于65株真菌中筛选到阳性菌株28株,其中产酶能力强的9株菌作为转化实验的备选菌株,以龙胆苦苷为底物进行转化,将转化粗提物进行TLC及HPLC分析,最终确定将C-5作为转化龙胆苦苷的菌株。根据菌株形态特征和核酸序列分析结果,初步鉴定菌株C-5为青霉属真菌。暂定名为Penicillium sp.C-5。  相似文献   

4.
5.
以濒危荒漠植物斧翅沙芥(Pugionium dolabratum)为对象,克隆脂质转运蛋白基因PdLTP3。运用Protparam、Multiple Sequence Alignment、MEGA、SignalP 4.1Server、TMHMM Server v.2.0、SOPMA、SWISS-MODEL等软件对其编码的氨基酸序列进行生物信息学分析,并检测干旱胁迫下PdLTP3基因的诱导表达情况。结果表明:从斧翅沙芥cDNA中克隆到PdLTP3基因全长,开放阅读框编码116个氨基酸,预测分子量为11.7kD,理论等电点8.91,平均亲水性系数0.569,是一个疏水蛋白质;同源比对显示其编码的蛋白质与拟南芥AtLTP3相似度较高;PdLTP3蛋白N端1~23个氨基酸序列为信号肽序列;二级和三级结构显示PdLTP3蛋白主要由α螺旋和无规则卷曲组成;干旱胁迫下PdLTP3基因表达量逐渐升高,5h后表达量约为胁迫处理前的48.6倍,说明其可能参与了斧翅沙芥干旱胁迫耐受。  相似文献   

6.
通过PCR结合变性梯度凝胶电泳(DGGE)技术对从南极长城站附近表层土壤样品中获得16SrDNA序列特征片段V3区序列进行分离。对其中的主要12条DGGE条带进行胶回收,获得的DNA片段经测序以及计算机比对分析发现,它们分别属于β、γ、δ-变形细菌(Proteobac-teria)、噬纤维菌-屈挠杆菌-拟杆菌(Cytophaga-Flexibacter-Bacteroides,CFB)群细菌、放线细菌(Actinobacteria)、蓝细菌属(Cyanobacteria)、酸杆菌属(Acidobacteria)和绿屈挠菌属(Chlo-roflexi)等系统分类群。南极表层土壤样品中的大部分16S rDNA序列与从其他土壤或沉积物样品中直接获得的序列相似性较高(93%-100%)。  相似文献   

7.
Gymnadenia conopsea,an alpine Orchidaceae plant,was one of the widely used Tibetan traditional medicines.In this study,we sequenced total 105 expressed sequence tags (ESTs) from a full-length cDNA expression library constructed by the Oligo-capping technique.The further bioinformatic analyses suggested that the 65 represented unique sequences showed high homology to previously identified genes in other plants:30 sequences matched to other uncharacterized expressed sequence tags (ESTs),and 10 sequences showed no good matches to available sequences in DNA databases.Gene ontology annotation by InterProScan indicated that many of these cDNAs (7 percent) have no known molecular functions and may be unique to G.conopsea.Fifty-five ESTs with matched proteins were involved in a series of diverse functions,in which molecular function such as "binding" (42.9 percent) and "catalytic activity" (25.0 percent) were the most frequent functions of the cDNAs.This cDNA library provided a critical basis for further investigation of functional genes expression under cold stress in this alpine species.In addition,13 ESTs-based polymerase chain reaction (PCR) primers were designed and can also be used for genotypic identification and for the genetic diversity analysis of G.conopsea and its closely related species.  相似文献   

8.
用于RAPD分析的沙拐枣DNA提取方法   总被引:14,自引:6,他引:8  
DNA分子标记技术是检测遗传差异的一种新技术,特别是RAPD技术,应用更为广泛。DNA质量是保证RAPD分析成功的关键。本文用改进的SDS法对沙拐枣属植物种子的总DNA进行了提取。琼脂糖凝胶电泳结果表明:得到的DNA片段大小在20kb以上;通过PCR扩增实验,证明用此方法提取的DNA,可直接用于随机扩增的DNA多态性(RAPD)遗传标记。  相似文献   

9.
从北极斯瓦尔巴群岛冰川流域的上、中、下游采集了5种代表性土壤和冰川融水样品(MT、AT、TN、BT、MS),采用常规LB和严格无氮培养基低温培养方法,对其中的可培养细菌和潜在固氮细菌进行了总菌计数以及菌株的富集、分离与纯化。经菌落菌体形态观察、16S r RNA基因初步测序排重、固氮基因nif H的PCR检测,分别选取48株可培养分离细菌和31株潜在固氮菌进行了基于16S r RNA基因全序列测定的系统发育分析。目的是初步了解该特殊生境中可培养细菌(包括潜在固氮菌)的多样性及其分布状况,为进一步研究它们在此生境中的生态功能奠定基础。实验结果表明,可培养细菌在5个采集样品中的总菌数为BTTNATMT=MS,严格无氮培养基分离菌总菌数为BTATMTTNMS。48株可培养代表菌分属于Proteobacteria、Actinobacteria、Bacteroidetes、Firmicutes 4个门的7纲、14属之中,它们在5个样品中的多样性程度为BT(6纲、6属)MT(4纲、6属)AT、MS(均为3纲、3属)TN(2纲、3属),优势菌群主要分布在γ-proteobacteria纲的Pseudomonas属(20.8%,集中在AT、BT、MS样品中)和β-proteobacteria纲的Janthinobacterium属(35.4%,集中在MS样品中),表现出一定程度的系统发育多样性。潜在固氮菌实验结果表明,能在严格无氮培养基上生长且可检测到nif H的31株代表菌具有极为单一的系统发育多样性。其中90.32%菌株集中在γ-proteobacteria纲的Pseudomonas属,仅在MT和MS样品中有3株菌分布在α-proteobacteria纲的Rhizobium属、β-proteobacteria纲的Janthinobacterium属和Actinobacteria纲的Arthrobacter属,它们与分离自其他极端环境的相关标准菌株具有高度16S r RNA基因同源性。另外,本实验还发现有8株菌的16S r RNA基因序列与相关标准菌株的相似度低于98.50%,推测它们具有成为该特殊生境下新种的潜质。  相似文献   

10.
耐旱苔藓植物DNA提取及优化RAPD、 ISSR反应体系的建立   总被引:1,自引:0,他引:1  
张道远  张元明  曹同 《中国沙漠》2006,26(5):826-831
 耐旱苔藓植物常常单个个体矮小、生物量低,如何从细小的单个个体中有效提取总DNA是进一步开展居群遗传多样性研究的关键。本研究以古尔班通古特沙漠广泛分布的刺叶墙藓(Tortula desertorum)为对象,使用快速提取法、2×CTAB法及DNeasy plant mini kit试剂盒提取法等3种方法对刺叶墙藓单个个体的总DNA进行提取。结果表明,2×CTAB法提取的DNA纯度高,凝胶电泳显示无明显降解现象,适宜作为PCR扩增的模板。利用所提取的单个个体DNA为模板,建立了优化的RAPD、ISSR反应体系。  相似文献   

11.
锦鸡儿属植物的遗传多样性及其种间关系   总被引:10,自引:3,他引:7  
利用RAPD技术,对甘肃省自然分布的11种锦鸡儿属(CaraganaFabr.)植物的遗传多样性及其种间关系进行了研究。从34个10碱基随机引物中筛选出能产生稳定多态性标记的引物11个,共扩增出128个位点,主要集中在200~2000bp之间,其中104个表现出多态性,多态性比率高达81.25%;根据POPGENE32软件计算的Shannon多样性指数与Nei指数也较高,分别为0.3889和0.2517。说明锦鸡儿属植物具有丰富的遗传多样性,对环境变化的适应能力较强。应用UPGMA法进行聚类分析,结果表明,RAPD分析所反映的种间遗传关系与传统的形态学分类结果基本一致,但也有个别种的从属地位稍有变化。这说明还需要将分子标记的研究结果与形态学和其他方法结合起来进一步进行研究。  相似文献   

12.
Here, we describe a technique that allows the genetic linage analysis of 16S rRNA genes in bacteria observed under a microscope. The technique includes the isolation of microbial cells using a laser microdissection microscope, lysis of the cells, and amplification of the 16S rRNA genes in the isolated cells without interference by bacterial DNA contamination from the experimental environment or reagents. Using this technique, we successfully determined 15 16S rRNA gene sequences in cells isolated from an Antarctic iceberg. These sequences showed 94%–100% identity to their closest strains, which included bacteria that occur in aqueous, marine, and soil environments.  相似文献   

13.
Chinese medaka(Oryzias sinensis),widely distributed in the plateau lakes,Yunnan Province,which was the indicative specie for the habitat quality of riparian zone in the wetland ecosystems and also was the key specie in the lake ecosystems,and also was one of the fishes who suffered the severe invasion of alien species.There,we investigated that the genetic diversity and population structure of the wild Chinese medaka by using inter-simple sequence repeat(ISSR)analysis.The samples(20 individuals in total)of Chinese medaka in three populations in the drainage area of Dian Lake were obtained.The 7 ISSR primers were picked out to assess the genetic diversity.The ISSR primers combinations revealed 82.69%polymorphism,among 52 genetic loci amplified.The sum of effective number of alleles and observed number of alleles were 1.492 4 and 1.826 9,respectively.The Shannon genetic diversity and Nei genetic diversity indexes of all the populations were 0.434 5 and 0.289 7,respectively.The gene differentiation(GST)was 0.198 9,and the gene flow(NM)was 2.013 2.The AMOVA demonstrated that only 6.65%of genetic variance came from inter-population,and 93.35%of genetic variance came from intra-population.Phylogenetic tree showed that 20 individuals from 3 populations were clustering mixed each other.Also,the principal coordinate analysis(PCA)supported the cluster analysis results.These results indicated that the genetic diversity of Oryzias sinensis was at the middle level,and there was no obvious genetic differentiation among the three populations of Oryzias sinensis.  相似文献   

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