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1.
Amino acid sequences of α-skeletal muscle actin isoforms in two species of rattail fish, Coryphaenoides acrolepis and Coryphaenoides cinereus 总被引：1，自引：1，他引：0
Takami Morita 《Fisheries Science》2000,66(6):1150-1157
SUMMARY: The cDNA clones of α-skeletal actins were isolated from the skeletal muscle of two species of rattail fish, Coryphaenoides acrolepis and Coryphaenoides cinereus . The complete nucleotide sequences of the cDNA and their deduced amino acid sequences were determined. Each of the two species had two α-skeletal actin cDNA. The nucleotide sequences of the coding region of the two α-skeletal actin isoform genes within each species had 92.0 and 91.8% homology. From the cDNA sequences of the four α-skeletal actin isoforms in the two species, amino acid sequences of 377 amino acid residues were deduced. It was predicted that the two N-terminal amino acid residues of each protein are processed after translation. The amino acid sequences of α-skeletal actin 1 in the two Coryphaenoides species were identical, as were the amino acid sequences of α-skeletal actin 2 in the two species. The amino acid sequences of the two α-actin isoforms, α-skeletal actin 1 and α-skeletal actin 2, differed by only a single amino acid, Ala/Ser at the 155th position. Northern blot analysis showed that a similar amount of each of the two α-actin isoform mRNA was expressed in the skeletal muscle of the two Coryphaenoides species. 相似文献
2.
采用同源克隆和RT-PCR技术对合浦珠母贝(Pinctada fucata)热休克蛋白hsp70基因进行了克隆和表达分析。获得cDNA全长序列2 365 bp,其中3’非编码区域(UTR)为318 bp,5’UTR为88 bp,开放阅读框(ORF)为1 959 bp,编码652个氨基酸,分子量约为71.39 kD,理论等电点为5.22,并含有3个HSP70家族的签名序列IDLGTTYS、DLGGGTFD和EEVD。同源性分析表明,合浦珠母贝HSP70的氨基酸序列与太平洋牡蛎(Crassostrea gigas)等双壳贝类的相似性高达86%以上,基于氨基酸序列的聚类分析表明,合浦珠母贝与牡蛎属种类亲缘关系最近。高温、高盐刺激后,半定量RT-PCR检测发现hsp70基因的表达明显增加,高温刺激的表达量高于高盐刺激,高温刺激组不同组织的表达量由大到小依次为鳃、消化腺、外套膜、肌肉、性腺,高盐刺激组不同组织的表达量由大到小依次为鳃、外套膜、肌肉、消化腺、性腺,表明HSP70参与了机体对刺激的应答过程。该基因的克隆为进一步深入研究合浦珠母贝的抗逆机理及其遗传改良奠定了重要基础。 相似文献
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为了解长牡蛎MITF基因的表达及其与壳色的关联,验证了长牡蛎中的4个MITF基因,对长牡蛎MITF氨基酸序列进行了序列分析和多序列比对,分析了长牡蛎幼体发育各时期的转录组,采用荧光定量PCR的方法研究长牡蛎各组织及黑壳、白壳牡蛎特定部位mRNA表达情况。4个MITF基因中有3个基因可能为假基因,有表达的长牡蛎MITF基因共编码448个氨基酸,为亲水性不稳定蛋白,含有N端结构域(MITFTFEBC3N superfamily)和高度保守的功能性结构域HLH结构域。转录组分析发现MITF在长牡蛎个体发育的各个时期均有表达,在稚贝期达到最高。组织表达结果显示MITF在外套膜中的表达水平显著高于其他组织。MITF在黑壳牡蛎闭壳肌中的表达量显著低于白壳牡蛎,而在黑壳牡蛎外套膜中的表达量高于白壳牡蛎,但不显著。在黑壳、白壳牡蛎外套膜边缘的表达量都显著高于内侧。研究表明,长牡蛎MITF基因可能在牡蛎壳形成早期就参与了黑色素的生成调控和个体的生长发育,可调控牡蛎酪氨酸酶Tyr2基因,参与外套膜和贝壳中黑色素的形成。本研究为进一步研究牡蛎壳色形成机制奠定了基础。 相似文献
4.
ABSTRACT: It has been reported that the amino acid sequences of striated and catch muscle myosin heavy chains from two scallop species ( Argopecten irradians and Placopecten magellanicus ) are almost identical, but that the ATPase activities between these myosins vary several-fold. These myosin sequences have been useful for identifying the region that modulates the ATPase activity of scallop myosin. In the present study, a cDNA encoding a myosin heavy chain was isolated from the mantle tissue of scallop Patinopecten yessoensis . The cDNA is composed of 6067 base pairs (bp) including an open-reading frame of 5841 p, which encodes an amino acid sequence of 1947 residues. The deduced amino acid sequence of P. yessoensis mantle myosin had a high identity of 90%, 92%, and 91% to P. magellanicus , A. irradians , and Pecten maximus striated muscle myosins, respectively. Interestingly, while the deduced amino acid sequences of around adenosine triphosphate-binding and actin-binding sites of the mantle myosin are homologous to those of A. irradians striated muscle myosin, the subfragment 2 hinge region and the non-helical tail region are similar to those of catch muscle myosin. 相似文献
5.
采用实验生态学方法,研究了盐度骤降(盐度分别为10、20,自然海水为对照组)对近江牡蛎(Crassostrea ariakensis)和长牡蛎(Crassostrea gigas)生理代谢的影响。结果显示,盐度骤降对长牡蛎和近江牡蛎的耗氧率(OR)、排氨率(NR)、排粪率(FER)均有显著影响(P<0.05),且2种牡蛎对盐度骤降的响应存在差异。长牡蛎的耗氧率、排氨率、摄食率(IR)和排粪率均在自然海水中最高,低盐对长牡蛎产生不同程度的胁迫。而近江牡蛎在盐度为20时的上述指标显著高于其他盐度处理组。通过测定不同规格牡蛎的代谢率,建立2种牡蛎在不同盐度下的代谢率与规格(湿重)的回归方程,该方程符合幂指数:Y=aWb。通过归一化处理,比较湿重在20 g时,2种牡蛎的能量分配差异。盐度为20时,近江牡蛎的呼吸代谢较强(呼吸能占16.1%),而长牡蛎在自然海水中呼吸代谢较强(呼吸能占7.8%),2种牡蛎的呼吸代谢均在盐度为10时较低,其中,长牡蛎呼吸能仅占0.7%。研究表明,盐度骤降对长牡蛎的影响大于近江牡蛎,小幅度的盐度骤降对近江牡蛎的能量代谢有一定的促进作用。本研究从能量代谢角度,探讨了近江牡蛎和长牡蛎对盐度骤变的响应,可为牡蛎礁修复工作和养殖生产过程中的品种选择提供参考。 相似文献
6.
长牡蛎壳金选育群体性腺发育与营养成分的周年变化 总被引：2，自引：2，他引：0
为了评估长牡蛎(Crassostrea gigas)壳金选育群体的繁殖周期及营养价值,自2013年11月至2014年10月,对山东省刘公岛海区长牡蛎第三代壳金选育群体的性腺发育、营养成分(总蛋白、总脂肪、糖原)季节变化及其与环境因子的关系进行了研究,并比较了与普通养殖群体营养成分的差异。结果表明,长牡蛎壳金选育群体雌雄比例接近1︰1,性腺同步发育;配子发生开始于2月,全年只有1个产卵季节(6―8月),7月为两性配子排放高峰期。配子成熟及排放期间,水温处于较高水平,食物充足,可为性腺发育提供能量,也有利于幼体的存活和生长。卵母细胞直径从配子发生开始逐渐增大,并在产卵前达到最大值(36.88μm),产卵后则降低。营养成分分析表明,脂肪含量在性腺–内脏团中随配子发育积累储存,产卵后显著降低;糖原在性腺发育到4月时开始下降,为配子的发育提供能量;蛋白质在配子发育过程中大量合成,与繁殖活动存在密切联系。长牡蛎壳金选育群体外套膜的总蛋白含量显著高于普通养殖群体(P0.05),糖原和总脂肪含量显著低于普通养殖群体(P0.05),其他组织的一般营养成分未出现显著差异。研究表明,长牡蛎在壳金群体选育过程中营养成分已表现出分化,这为长牡蛎壳色选育提供了重要参考依据。 相似文献
7.
蛋白酶抑制因子是极其多样的蛋白质或多肽,能抑制蛋白酶的水解活性。研究发现蛋白酶抑制因子能通过抑制病原蛋白酶活性,从而抑制病原的入侵。I84蛋白酶抑制因子家族是MEROPS数据库中新增的一个家族,其部分成员在免疫防御过程中的作用得到了一定的揭示。为探究I84蛋白酶抑制因子家族在长牡蛎中的分布和功能状况,实验鉴定了长牡蛎中23个潜在的I84家族基因,根据系统进化树分析,挑选了5个同源基因进行时空表达和功能分析。首先,利用克隆技术验证了长牡蛎中5个I84家族同源基因CgSi3、CgSi5、CgSi6、CgSi16和CgSi19可表达性。结果显示,5个基因在外套膜、闭壳肌、性腺、血淋巴细胞、肝胰腺和鳃等6个组织中均表达,但肝胰腺中表达量显著高于其他组织。同时,5个基因在长牡蛎幼体不同发育阶段表达模式不同,其中各基因在受精卵时期均不表达,CgSi3表达量则在眼点幼虫期显著上升后下降,而CgSi6在壳顶幼虫期开始表达后,表达量随长牡蛎发育而持续增加。另外,对牡蛎进行病原相关模式分子(PAMPs)注射后,5个基因也表现出不同的表达模式,其中LPS和PGN注射后CgSi6表达量变化明显,而poly (... 相似文献
8.
5种经济头足类水产品加工工艺性质的研究 总被引：1，自引：0，他引：1
以新鲜的枪乌贼、太平洋褶柔鱼、短蛸、冷冻的柔鱼和茎柔鱼为原料,分别测定其质量组成及其一般化学成分,并比较了柔鱼和茎柔鱼的热加工特性。研究发现,5种头足类水产品的可食部分均超过70%,水分含量超过78%,粗蛋白含量为58%~83%,粗脂肪含量为4%~19%。在2种不同的热处理方式下,柔鱼和茎柔鱼外套膜沿体轴方向的收缩均大于环绕体轴方向的收缩,在蒸煮过程中,柔鱼的质量损失和水分流失均在5 min时达到最大,茎柔鱼在6 min时达到最大,在烘烤的过程中,2种柔鱼的质量损失随着水分流失的增加而增加。 相似文献
9.
肌细胞增强因子(Myocyte enhancer factor-2,MEF2)是一种肌肉特异性转录因子,对脊椎动物肌纤维的形成和发育具有重要的调控作用。为了研究MEF2对华贵栉孔扇贝(Chlamys nobilis)闭壳肌肌纤维形成和发育的调控作用,实验克隆了华贵栉孔扇贝MEF2C和MEF2C-like c DNA序列,解析了其在不同组织和发育时期的表达调控规律,分析了盐度对其表达调控的影响。结果表明,MEF2C和MEF2C-like基因开放阅读框(Open Reading Frame,ORF)长为1 302 bp和1 158 bp,分别编码433和358个氨基酸。氨基酸序列比对分析显示MADS和MEF2结构域高度同源,系统进化分析显示华贵栉孔扇贝和长牡蛎具有较近的亲缘关系。在不同组织和发育时期,华贵栉孔扇贝MEF2C和MEF2C-lik mRNA具有相似的表达规律,均在D型幼虫期及闭壳肌中显著高表达。在不同日龄的华贵栉孔扇贝闭壳肌中,MEF2Cs mRNA在60和120日龄表达水平显著升高。MEF2Cs mRNA在盐度为28时表达量最高,说明可能在此盐度下更适合华贵栉孔扇贝生长。 相似文献
10.
文蛤肠、外套膜和肝胰脏组织 cDNA 文库构建及其 ESTs 序列分析 总被引：3，自引：0，他引：3
利用SMART cDNA文库构建试剂盒构建了文蛤(Meretrix meretrix)肠、外套膜和肝胰脏组织的cDNA文库。经测定原始文库滴度分别为2.10×106、1.70×106和1.60×106,重组率高于95%,肠组织文库插入片段长度均大于1000bp,外套膜和肝胰脏组织文库的插入片段长度大于1kb的占87.5%,文库质量符合标准要求。随机选取文库的3168个克隆进行5′端测序,获得高质量表达序列标签(Expressed Sequence Tags,ESTs)3029个(肠:1005,外套膜:1019,肝胰脏:1005),测序成功率95.58%。经质量控制和拼接得到1796个单基因簇(Unigene),其中306个叠联群(Contigs),1490个单一序列(Singletons)。通过Blastx搜索比对、查询和注释分析,共得到已知基因696个(肠:216,外套膜:235个;肝胰脏:245个),占总数38.75%。在1796个单基因簇(Unigene)发现微卫星序列55条,这些存在微卫星位点的序列占整个ESTs数据库的3.1%。 相似文献
11.
Masaya TAKAGI Takeshi KIKKO Masatomi HOSOI Isao HAYASHI Haruhiko TOYOHARA 《Fisheries Science》2004,70(4):682-687
ABSTRACT: We have cloned a cDNA encoding the matrix metalloproteinase (MMP) from oyster Crassostrea gigas . The clone contains a 1797 bp open reading frame encoding a protein of 599 amino acids. Oyster MMP (Cg-MMP) has a transmembrane domain at its C-terminal and a furin/prohormone convertase cleavage site at the end of a propeptide domain, which are commonly observed in membrane-type MMPs (MT-MMPs). This suggests that Cg-MMP is an MT-MMP. The deduced amino acid sequence of oyster MMP shares approximately 30% identity with human MT4-MMP and MMPs from fruit fly and hydra. Cg-MMP mRNA was detected in the gill, mantle and adductor muscle, and more intense signals in the northern blot analysis were recognized in the gill and adductor muscle. Similar tissue distribution was observed for tissue inhibitor of metalloproteinase (Cg-TIMP) in oyster. In response to hypoxic stress, the abundance of Cg-MMP mRNA was elevated in the gill, while that of Cg-TIMP mRNA remained almost constant. These findings suggest that promotion of collagen metabolism may be implicated in the hypoxic adaptation in oyster. 相似文献
12.
ABSTRACT: Genetic differentiation and relationships between Crassostrea plicatula and Crassostrea gigas populations from China were studied by means of the microsatellite technique. Seven loci were used to screen five populations each collected from C. plicatula and C. gigas . All loci showed high polymorphism for all populations, as observed in average number of alleles per locus (19.1–28.1), and average expected heterozygosity (0.891–0.954). Significant departures from Hardy–Weinberg equilibrium due to heterozygote deficiency were observed over most populations at each locus and were best explained by null alleles. F ST values showed significant genetic differentiation between C. plicatula and C. gigas populations. According to the neighbor-joining tree constructed on the basis of the genetic distance ( D A ), the ten populations fell into two distinct groups ( C. plicatula and C. gigas groups), and the results of principal coordinate analysis and assignment tests also supported the neighbor-joining clustering. The outcomes presented here suggested that the microsatellite markers have great potential for differentiating C. plicatula from C. gigas populations. The information obtained in this study has important implications for the suitable management and conservation of these genetic resources in China. 相似文献
13.
Cristian Gallardo‐Escárate Valentina Valenzuela‐Muñoz Gustavo Núñez‐Acuña Pilar Haye 《Aquaculture Research》2015,46(5):1175-1187
The main objective of this research was to identify single‐nucleotide polymorphisms (SNPs) from an Expressed Sequences Tags (EST) data set generated by 454 pyrosequencing in the soft clam Mesodesma donacium. A total of 180 159 ESTs were yielded from a M. donacium cDNA library. De novo assembly was performed using stringent calling parameters, producing 10 178 contigs and 41 765 singletons. Here, a total of 2594 SNPs were discovered related to 613 consensus sequences, achieving a frequency of 1 SNPs per 260 bp. SNP variants showed that A/G, A/T and C/T were the most abundant among the identified polymorphisms. We validated a total of 12 SNPs loci by HRMA for annotated genes such as heat shock protein‐70 and the translation elongation factor 1‐alpha. The Gene Ontology analysis regarding molecular function level revealed that sequences with SNPs were mainly classified to protein and nucleotide binding, as well hydrolase activity, ion binding and oxidoreductase activity. Further, biological processes like cellular and metabolic process, biogenesis, localization and biological regulation were highly annotated. The most expressed genes were related to the mitochondrial electron transport chain, senescence‐associated protein, ubiquitin and actin. Interestingly, some relevant genes related to immune response and biomineralization showed a high abundance, such as tumor necrosis factor (TNF)‐alpha‐receptor‐like protein, serine protease inhibitor, heat shock protein, aragonite‐binding protein and ferritin. This study contributes to relevant genes associated with functional polymorphisms and gives an overview for future genetic investigations. 相似文献
14.
Yukihiro Kitade Michiko Nakamura Hirotoshi Endo Satoru Fukuda Kazuyoshi Kuwano Naotsune Saga 《Fisheries Science》2006,72(3):639-645
ABSTRACT: A cDNA ( PyARP4 ) containing an open reading frame for a protein of 573 amino acids was identified in the marine red alga Porphyra yezoensis . The conceptual PyARP4 protein exhibits significant similarity to actin-related protein (ARP) 4 in the terrestrial plant Arabidopsis . Comparison of the deduced amino acid sequence showed moderate sequence identity (30%) to a conventional actin in P. yezoensis , as seen in comparisons between ARP and conventional actins of other organisms. A putative bipartite nuclear localization signal and an actin motif were found within the PyARP4 amino acid sequence. In a phylogenetic analysis, the PyARP4 was found to cluster with the ARP4 of other organisms. The expression level of PyARP4 did not change significantly among four developmental stages of life cycle and was lower than that of a conventional actin. This cDNA therefore may serve as a useful internal standard in gene expression analyses of differentially expressed genes in P. yezoensis . 相似文献
15.
16.
大菱鲆脾脏cDNA文库构建、EST序列分析与免疫抗病相关基因的筛选 总被引：1，自引：0，他引：1
以大菱鲆脾脏为材料,构建cDNA文库,通过对文库克隆的序列测定和初步生物信息学分析,共获得3 656个大菱鲆脾脏表达序列标签（EST）。经与GenBank数据库的序列比对后发现,1 891个EST代表了149个已知基因,其余1 765个EST为未知序列。这149个基因大致可分为6类：9个为细胞结构类（6.0%）;26个为代谢类（17.5%）;45个为细胞防御/免疫类（30.2%）;43个为基因/蛋白质表达类（28.9%）;16个为细胞信号转导/细胞交流类（10.7%）;10个无法明确分类（6.7%）。鉴定的与免疫抗病相关重要基因主要有：CC/CXC趋化因子;主要组织相容性复合体（MHC）class I α;C3补体;天然杀伤细胞增强因子（NKEF）;胸腺素β-4/β-12;干扰素诱导蛋白Gig1;白介素8受体;STAT 4;热激蛋白70/90等。 相似文献
17.
秘鲁外海茎柔鱼角质颚的形态特征及其与个体大小和日龄的关系 总被引：2，自引：0，他引：2
角质颚是头足类重要的硬组织,被用于估算其个体大小及其生物量等。根据2009~2014年中国鱿钓船在秘鲁外海(79°22'W~84°30'W、10°00'S~18°16'S)采集的茎柔鱼(Dosidicus gigas)样本,提取出1 346ind茎柔鱼的角质颚,并对角质颚的形态进行了分析,建立了角质颚的形态特征参数与胴长、体重和日龄之间的关系。结果显示,雌性个体角质颚各形态参数的平均值均大于雄性个体,在角质颚12个形态参数中,上喙长(UHL)、上脊突长(UCL)、上侧壁长(ULWL)、下脊突长(LCL)、下侧壁长(LLWL)、下翼长(LWL)与胴长、体重、日龄之间的相关系数较高。胴长与角质颚各形态参数的关系在雌、雄个体间的差异性均显著(P0.05),体重、年龄与角质颚各形态参数的关系在雌、雄个体间的差异性均不显著(P0.05)。角质颚各形态参数与胴长、日龄之间呈线性关系;除上侧壁长外,角质颚的其它形态参数与体重之间呈幂指数关系。主成分分析认为,第一主成分解释上、下角质颚形态的贡献率分别为95.69%和95.11%,在上角质颚和下角质颚的形态参数中,载荷系数最大的形态参数分别为上脊突长和下侧壁长。研究认为,上述关系式的建立可以用于茎柔鱼的个体大小、生物量和日龄的估算。 相似文献
18.
广东沿海贝类4种重金属含量分析和评价 总被引：8，自引：0，他引：8
根据2007年6～9月对广东沿海4种贝类养殖区90个样品的检测结果，研究了贝类体内重金属的含量和累积，对贝类的质量安全进行评价。结果表明：（1）菲律宾蛤仔Ruditapes philippinarum体内含量大小顺序为铜（Cu）〉镉（Cd）〉铬（Cr）〉铅（Pb），太平洋牡蛎Crassostreagigas和近江牡蛎C．rivularis的大小顺序都是Cu〉Cd〉Pb〉Cr，而翡翠贻贝Pernaviridis的大小顺序则是Cu〉Pb〉Cd〉Cr；（2）菲律宾蛤仔、太平洋牡蛎和翡翠贻贝未受重金属污染，但部分近江牡蛎体中Cd和Cu含量已超过中国农业行业标准“无公害食品：水产品有毒有害物质限量”标准，应该引起重视。 相似文献
19.
栉孔扇贝wnt4基因cDNA克隆及表达分析 总被引：2，自引：0，他引：2
由栉孔扇贝(Chlamys farreri)转录组数据库获得wnt4基因的一段表达序列标签(EST)序列,利用SMART-RACE技术克隆了wnt4基因cDNA全长序列,该序列长1 239 bp,其中开放阅读框为1 068 bp,可以编码355个氨基酸,推导的氨基酸序列含有WNT家族特有序列,且与沙蚕(Platynereis dumerilii)、海胆(Heliocidariserythrogramma)、人(Homo sapiens)等物种WNT4同源性都在60%以上。半定量RT-PCR结果显示,除肾脏外,wnt4基因在栉孔扇贝的精巢、卵巢、闭壳肌、肝胰腺、鳃、外套膜组织中均有表达,但表达量较低。定量RT-PCR结果表明:wnt4基因在成熟期的精卵巢中表达量最高,增殖期和休止期表达量次之,生长期表达量最低;整个生殖周期中精巢表达量高于卵巢。该基因在栉孔扇贝多个组织中的表达特性,暗示其参与了多样的生物学过程;在性腺中的表达特征表明其可能参与两性性腺的发育并在生殖细胞成熟过程中发挥作用。 相似文献
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