文蛤亮氨酸氨肽酶LAP3的基因克隆及其在幼体不同发育时期、成体不同组织的表达分析     

阮文斌  董迎辉  高晓艳  刘晨珊  林德海  林志华 《海洋学报》2017,39(2):96-104

为探索贝类LAP3基因的结构和功能特征以及在生长发育过程中的作用,利用cDNA末端快速扩增（RACE）法获得文蛤LAP3（Mm-LAP3）基因的cDNA全长序列,并对其生物信息学、组织及发育时期表达特征进行了分析。结果显示,Mm-LAP3基因cDNA全长2 037 bp,ORF区1 254 bp,编码417个氨基酸;Mm-LAP3蛋白由Peptidase_M17超家族序列的N-端结构域和Peptidase_M17催化结构域组成。荧光实时定量PCR结果表明,Mm-LAP3基因在文蛤成体6个组织和幼体10个发育时期中均有表达,其中在斧足中的表达量显著高于其他组织（P<0.05）,这与斧足蛋白含量丰富、代谢旺盛相关;在幼体10个发育时期中的表达量呈现逐渐升高的趋势,在D形幼虫时期达到最高,随后又有所降低,推测Mm-LAP3基因在早期发育时期参与了某些组织器官的形成。  相似文献   

红笛鲷(Lutjanus sanguineus)CD4和CD4-2基因克隆与诱导表达     

黄郁葱  丁燏  梁秀全  蔡双虎  吴灶和  简纪常 《海洋与湖沼》2017,48(3):589-599

应用RT-PCR和RACE-PCR技术克隆了红笛鲷(Lutjanus sanguineus)CD4和CD4-2基因,并分析了它们在健康鱼不同组织的表达分布及免疫刺激物诱导后的表达变化。CD4基因c DNA序列全长2216bp,包含180bp的5′UTR(untranslated regions)、605bp的3′UTR和1431bp的开放阅读框(open reading frame,ORF),编码476个氨基酸;红笛鲷CD4-2基因c DNA序列全长为1520bp,包含62bp的5′UTR、525bp的3′UTR和933bp的ORF,编码310个氨基酸。CD4分子由信号肽、4个免疫球蛋白样结构域(D1—D4)构成的胞外区、跨膜区和胞浆区组成,CD4-2分子由信号肽、2个免疫球蛋白样结构域(D1—D2)构成的胞外区、跨膜区和胞浆区组成。荧光定量PCR(Real Time Quantitative PCR,q PCR)分析显示红笛鲷CD4和CD4-2基因在健康鱼的胸腺中表达量最高,其次是中肾、鳃、皮肤、脾、头肾和肠。红笛鲷头肾淋巴细胞体外经脂多糖(Lipopolysaccharide,LPS)和刀豆蛋白A(Concanavalin A,Con A)刺激12h后,CD4和CD4-2表达量显著上调(P0.05)。哈维氏弧菌疫苗免疫24h后鳃、头肾、脾脏和肠的表达量显著上升(P0.05)。研究结果为进一步研究鱼类CD4和CD4-2在抗菌免疫反应中的作用提供了基础资料。  相似文献   

Prevalence of Bacillus sp. among the biofilm forming community on Ti surface in marine environment     

PRIYA Chokkalingam  ARAVIND Ganessin  THILAGARAJ Wilson Richard 《海洋学报(英文版)》2017,36(6):89-94

Prevalence of bacterial species involved in biomineralization of manganese on titanium (Ti) surfaces in marine environment was revealed in this research work. This study involves one year sea water exposure of Ti and their periodical biofilm characterization was carried out to quantify the manganese oxidizing bacterial (MOB) presence in the biofilm formed on titanium surfaces. The total viable count study of Ti coupons exposed to sea water for one year resulted in 60% of the MOB in overall biofilm population. The biochemical characterization of MOB isolates were performed for the genus level identification of the seven bacterial isolates. Further, the seven strains were subjected to 16S rRNA gene sequencing. Evolutionary analysis was performed using MEGA 7 to obtain closely related strains within the groups. The manganese oxidizing ability of the bacterial isolates were determined with Leucoberbelin Blue Assay (LBB) and Atomic Absorption Spectroscopy studies (AAS). The results show that among the isolated marine MOB species, Bacillus sp. and Leptothrix sp. have the maximum Mn oxidizing property. The microtitre plate assay was performed to determine the biofilm forming ability of the isolated marine MOB species. All the results have confirmed the prevalence of Bacillus sp. among the biofilm colonizers on Ti surfaces when exposed in sea water.  相似文献   

施氏鲟Asnanos1基因序列分析及其在雌雄不同组织中表达     

肖懿哲  朱友芳  孙玉龙  张鑫  张子平  王艺磊 《海洋科学》2017,41(4):17-23

本研究首先从施氏鲟(Acipenser schrenckii)性腺转录组中获得nanos1(Asnanos1)基因全长c DNA,并对其序列的准确性和特征分别进行PCR验证和生物信息学分析。进一步利用荧光定量RT-PCR技术检测Asnanos1基因在雌、雄施氏鲟的性腺、心脏、鳃、脑、肾、肝脏、脾脏等组织中的表达量。利用荧光定量RT-PCR技术检测Asnanos1基因在雌、雄施氏鲟的性腺、心脏、鳃、脑、肾、肝脏、脾脏等组织中的表达量。结果显示:Asnanos1的c DNA全长为1 389 bp,其中,5′非编码区(UTR)为414 bp,3′UTR为279 bp,ORF为696 bp。该基因共编码231个氨基酸。Asnanos1基因在施氏鲟除心脏以外的各组织中均有表达,在雌、雄鱼的鳃、脑、肾、肝脏、脾脏等组织中表达量无显著差异,但在卵巢中的表达量显著高于其他各组织(P0.05)。所得到的结果可为人工培育全雌施氏鲟苗种提供一些基础资料。  相似文献   

西南印度洋真光层海水中固氮细菌多样性     

谢尚微  杨俊毅  张东声  王春生 《海洋学研究》2015,33(3):54-61

本文通过构建克隆文库和基因测序的方法研究了西南印度洋真光层海水中固氮细菌nifH基因的多样性。从构建的2个nifH基因克隆文库中共获得76条有效序列,其中46条来自CTD13-30 m文库,分属10个OTUs;30条来自CTD13-125 m文库,分属8个OTUs。系统发育分析结果显示,研究站位nifH基因序列主要分布于Cluster I和Cluster III两个分支,其中Cluster I中包含蓝细菌和变形菌两个分支,蓝细菌以Group B为优势类群,并未获得束毛藻和Group A的nifH基因序列。此外还有少数nifH基因序列分布于Cluster II。总体来看,西南印度洋固氮生物基因与大西洋的亲缘关系更近;固氮生物的多样性较为丰富,受环境条件的影响,群落结构与其它热带、亚热带寡营养海域具有明显的不同。  相似文献   

客家文化景观基因特征 ——以湖南省炎陵县为例   总被引：1，自引：0，他引：1  

曹帅强  邓运员  杨载田  何清华  李敏 《热带地理》2014,34(6):831-841

以中国南方山丘区具有代表性的客家文化聚居区--炎陵县为例,采用客家文化景观基因的特征提取技术,从民居建筑、传统习俗、地方方言、宗法制度、民间信仰5个角度,提取了炎陵县客家文化景观基因的特征。研究结果表明：1）该县客家人以开放性“围堡式”的血缘宗族聚居模式,与畲、黎、瑶等少数民族通婚繁衍在山丘区的山谷地带;在“入乡随俗”谋求生存发展的环境中,某些方面已适应当地习俗,但仍保留了基本的客家精神及风俗;并以当地炎帝神农文化为主要文化参照,形成了独具炎陵县特色的客家文化。2）该县客家文化的景观基因在不同发生发展过程中有着衣、食、住、行、商等物质性方面的表达载体和地方语言、宗法制度、民间信仰、民情风俗、文化艺术等非物质性方面的表达形式。  相似文献   

3种典型荒漠灌木内生固氮菌及固氮酶基因nifH多样性分析   总被引：1，自引：0，他引：1  

徐正金  罗明  王卫霞  王纯利 《中国沙漠》2014,34(2):472-480

具有特殊生境的内生固氮菌,对改善植物营养、增强植物抗逆性及群落稳定性具有重要作用,是一类潜力巨大、尚待开发的微生物资源。以新疆3种典型荒漠灌木多枝柽柳（Tamarix ramosissima）、黑果枸杞（Lycium ruthenicum）、盐穗木（Halostachys caspica）为材料,从根和枝条组织中分离筛选获得137个内生固氮菌菌株,采用16S rDNA PCR-RFLP、16S rDNA序列测定、BOXAIR-PCR指纹图谱、nifH PCR-RFLP等方法分析其遗传多样性及系统发育关系。结果表明：分离菌株经16SrDNA PCR-RFLP酶切图谱聚类划分为9个遗传类型。序列测定和系统发育分析显示,代表菌株分属于拉恩氏菌属（Rahnella）、假单胞菌属（Pseudomonas）、泛菌属（Pantoea）、芽孢杆菌属（Bacillus）、寡养单胞菌属（Stenotrophomonas）和申氏杆菌属（Shinella）,其中Rahnella为优势种群。一个测定菌株的16SrDNA序列同源性低于97%,提示可能为一个潜在的新种。BOXAIR-PCR分析优势种群的基因组结构特征,获得12种指纹图谱类型,显示出该种群具有丰富的基因组多样性。nifH PCR-RFLP分析分离内生固氮菌菌株固氮酶基因nifH的分子多态性,将其划分为3种基因型,体现了nifH具有高度保守性,同时在不同固氮微生物种群和菌株间也存在一定的多样性。研究结果丰富了固氮微生物物种资源库和基因库,对于内生固氮菌资源的保护和利用奠定了科学基础。  相似文献   

Molecular Cloning of a Cytosolic G6PDH Gene from Populus Suaveolens and Its Expression to Improve the Cold Resistance of Tobacco Plants     

LIN Yuan-zhen  GUO Hai  ZHANG Zhi-yi  LIN Shan-zhi 《冰川冻土》2009,31(6)

Glucose-6-phosphate dehydrogenase (G6PDH, EC 1.1.1.49) is the first and main regulated enzyme of oxidative pentose phosphate pathway (OPPP), catalyzing the conversion of glucose-6-phosphate to 6-phospho-gluconolactone and playing important roles in the growth and development of plants. It is preciously reported that the enhancement of freezing resistance of Populus suaveolens cuttings is clear related to the distinct increase in cytosolic G6PDH activity. Here, a 1697 bp cDNA fragment (PsG6PDH) is amplified by RT-PCR from cold-induced total RNA of the freezing-tolerant P. suaveolens . A sequence analysis showed that PsG6PDH coding region had 1 530 bp and encoded 510 predicted amino acid residues. Genomic Southern analysis revealed that the isoform is encoded by a few copies of the gene in the poplar genome. The cloned gene PsG6PDH is cloned into binary vector pBI121 and used to transform tobacco. PCR and Southern blotting results verified integration of this gene into the genome of tobacco. Moreover, cold treatment experiments and membrane defense enzyme activity analysis confirmed that overexpression of the PsG6PDH gene could enhance the tolerance to cold or frigid stresses in transgenic plants.  相似文献   

Phylogenetic diversity of planktonic bacteria in the Chukchi Borderland region in summer   总被引：1，自引：1，他引：0  

ZENG Yinxin  YU Yong  LI Huirong  HE Jianfeng  LEE Sang H  SUN Kun 《海洋学报(英文版)》2013,32(6):66-74

Planktonic bacteria are abundant in the Chukchi Borderland region. However, little is known about their diversity and the roles of various bacteria in the ocean. Seawater samples were collected from two stations K2S and K4S where sea ice was melting obviously. The analysis of water samples with fluorescence in situ hybridization (FISH) showed that DMSP-degrading bacteria accounted for 13% of the total bacteria at the station K2S. No aerobic anoxygenic phototrophic (AAP) bacteria were detected in both samples. The bacterial communities were characterized by two 16S rRNA gene clone libraries. Sequences fell into four major lineages of the domain Bacteria, including Proteobacteria (Alpha, Beta and Gamma subclasses), Bacteroidetes, Actinobacteria and Firmicutes. No significant difference was found between the two clone libraries. SAR11 and Rhodobacteraceae clades of Alphaproteobacteria and Pseudoalteromonas of Gammapro-teobacteria constituted three dominant fractions in the clone libraries. A total of 191 heterotrophic bacterial strains were isolated and 76% showed extracellular proteolytic activity. Phylogenetic analysis reveals that the isolates fell into Gammaproteobacteria, Bacteroidetes, Actinobacteria and Firmicutes. The most common genus in both the bacterial isolates and protease-producing bacteria was Pseudoalteromonas. UniFrac data showed suggestive differences in bacterial communities between the Chukchi Borderland and the northern Bering Sea.  相似文献   

凡纳滨对虾P23蛋白基因的筛选、表达和生物信息学分析     

李喜莲  张艳艳  辛静静  钱昭英  刘小林  相建海 《海洋学报》2013,35(4):162-167

克隆凡纳滨对虾极端体重个体的组织差异表达基因P23基因并进行生物信息学分析, 为进一步研究该基因的功能以及凡纳滨对虾的分子选育等研究提供信息。以凡纳滨对虾雌虾极端体重个体腹部肌肉为实验材料, 利用抑制消减杂交(SSH)技术构建极大体重雌性个体和极小体重雌性个体腹部肌肉组织正反向消减cDNA文库:正库(以极大体重个体为试验组, 以极小体重个体为驱动组, L-S)和反库(以极小体重个体为试验组, 以极大体重个体为驱动组, S-L)消减cDNA文库, 并采用实时定量技术分析P23基因的组织表达规律, 利用生物信息学对其功能和结构进行预测。以β-actin为看家基因检测两个文库的消减效率分别为210和25, 实时定量技术分析结果表明P23基因在体重的调节中起上调作用。P23基因编码区序列全长495bp, 编码165个氨基酸, GenBank登录号:JF806619。生物信息学分析发现P23蛋白部分序列含有强疏水性, 无跨膜螺旋结构, 两种信号肽预测都显示P23含有信号肽。  相似文献