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1.
The complete nucleotide sequence of the mitochondrial genome of long-finned squid Loligo edulis forma kensaki (L. edulis f. kensaki, Teuthida: Myopsida) was determined. It contains 13 protein-coding genes, 12SrRNA, 16SrRNA, and the 22 tRNA genes, and three long noncoding regions. We also determined the mitochondrial genome nucleotide sequence of seasonal brood samples of L. edulis f. budo collected from the western part of the Sea of Japan in the autumn to winter seasons, which showed morphologic discrepancies with L. edulis f. kensaki. Comparison of the two complete sets of mitochondrial DNA data showed that the similarity of the two sequences was as high as 99.9?%, with only eight nucleotide substitutions in coding regions. The complete mitochondrial genome of Loligo edulis f. kensaki from Nagasaki Pref. is 17360?bp and L. edulis f. budo from the Shimane Pref. is 17351?bp. The nucleotide sequences and haplotype patterns in ND1 suggest that L. edulis f. kensaki and L. edulis f. budo share a single genetic background. 相似文献
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Jixuan Yang Qingsong Tan Wenhuan Zhu Chen Chen Xufang Liang Lei Pan 《Fish physiology and biochemistry》2014,40(1):93-104
The solute carrier family 7A, member 7 gene encodes the light chain- y+L amino acid transporter-1 (y+LAT1) of the heterodimeric carrier responsible for cationic amino acid (CAA) transport across the basolateral membranes of epithelial cells in intestine and kidney. Rising attention has been given to y+LAT1 involved in CAA metabolic pathways and growth control. The molecular characterization and function analysis of y+LAT1 in grass carp (Ctenopharyngodon idellus) is currently unknown. In the present study, full-length cDNA (2,688 bp), which encodes y+LAT1 and contains a 5′-untranslated region (319 bp), an open reading frame (1,506 bp) and a 3′-untranslated region (863 bp), has been cloned from grass carp. Amino acid sequence of grass carp y+LAT1 contains 11 transmembrane domains and shows 95 %, 80 % and 75 % sequence similarity to zebra fish, amphibian and mammalian y+LAT1, respectively. The tissue distribution and expression regulation by fasting of y+LAT1 mRNA were analyzed using real-time PCR. Our results showed that y+LAT1 mRNA was highly expressed in midgut, foregut and spleen while weakly expressed in hindgut, kidney, gill, brain, heart, liver and muscle. Nutritional status significantly influenced y+LAT1 mRNA expression in fish tissues, such as down-regulation of y+LAT1 mRNA expression after fasting (14 days). 相似文献
3.
利用半滑舌鳎性腺转录组测序获得的StAR基因部分序列,设计RACE引物,克隆了半滑舌鳎StAR基因的cDNA序列,全长为1 294 bp,5'端UTR为132 bp,3'端UTR为310 bp,开放阅读框(ORF)为852 bp,共编码283个氨基酸。将半滑舌鳎StAR基因与其他物种StAR基因进行氨基酸同源性分析,结果显示,半滑舌鳎StAR与塞内加尔鳎、大口黑鲈、花鲈、金头鲷的同源性都达到了85%,与虹鳟、斜带石斑鱼及日本鳗鲡的同源性分别为81%、83%和76%。雌、雄鱼不同组织StAR基因的表达分析表明,StAR基因在雄鱼性腺中高表达,在雄鱼的肝脏、脑及心脏中表达量较低,而在雄鱼的其他组织中不表达;在雌鱼肠中不表达,在其他组织(卵巢、肝脏、脾脏、脑、垂体、肌肉、心脏、肾脏)中微量表达。荧光定量PCR分析不同组织与不同时期性腺表达谱表明,雄鱼性腺中StAR基因的表达量显著高于雌、雄鱼其他各组织(P0.05),提示StAR基因对雄鱼精巢发育起重要作用。雄鱼不同时期表达谱分析结果显示,StAR基因在66天前的精巢中不表达,在150天时表达量急剧增加,至2龄时表达量最高,3龄时表达量下降,说明该基因在精巢发育成熟过程中起重要作用。原位杂交结果显示,StAR基因主要在雄鱼精巢的精子细胞中表达,而在雌鱼的卵巢中不表达。研究表明,StAR基因在半滑舌鳎精巢发育中发挥作用,且可能在精子形成中发挥重要作用。 相似文献
4.
Tian Yuan Chuang Li Ji-Rui Gu Yue-Jun Fu Heng Xu 《Fish physiology and biochemistry》2013,39(4):829-835
An expressed sequence tag of grass carp leukocyte cell–derived chemotaxin 2 (LECT2) gene was screened from an established intestinal cDNA library. Rapid amplification of cDNA ends gave rise to a full-length LECT2 cDNA (gcLECT2) with a complete open-reading frame of 474 bp, encoding 158 amino acids about 17.9 kDa. Homology search and sequence alignment showed that this deduced protein sequence shared a high identity with LECT2 from other vertebrates. Western blotting indicated immunological cross-reactivity occurs between grass carp and human LECT2 protein. This gcLECT2 genomic sequence is 1,868 bp in size, which consists of five exons and four introns. Real-time quantitative PCR analysis revealed that gcLECT2 gene is ubiquitously expressed in different tissues of healthy grass carp including brain, gut, liver, spleen, kidney, muscle and heart, while the expression levels were significantly increased in liver and spleen followed by Aeromonas salmonicida infection. 992 bp 5′-flanking region sequence was cloned and analyzed, where one CAAT box and one GC island were found. Our results showed that the LECT2 is suggested to be most possibly involved in the grass carp’s immune response. 相似文献
5.
Lin Zeng Bin Liu Chang-Wen Wu Ji-Lin Lei Mei-Ying Xu Ai-Yi Zhu Jian-She Zhang Wan-Shu Hong 《Fish physiology and biochemistry》2016,42(6):1595-1607
AMP-activated protein kinase (AMPK) is a highly conserved and multi-functional protein kinase that plays important roles in both intracellular energy balance and cellular stress response. In the present study, molecular characterization, tissue distribution and gene expression levels of the AMPK α1 and α2 genes from turbot (Scophthalmus maximus) under salinity stress are described. The complete coding regions of the AMPK α1 and α2 genes were isolated from turbot through degenerate primers in combination with RACE using muscle cDNA. The complete coding regions of AMPK α1 (1722 bp) and α2 (1674 bp) encoded 573 and 557 amino acids peptides, respectively. Multiple alignments, structural analysis and phylogenetic tree construction indicated that S. maximus AMPK α1 and α2 shared a high amino acid identity with other species, especially fish. AMPK α1 and α2 genes could be detected in all tested tissues, indicating that they are constitutively expressed. Salinity challenges significantly altered the gene expression levels of AMPK α1 and α2 mRNA in a salinity- and time-dependent manners in S. maximus gill tissues, suggesting that AMPK α1 and α2 played important roles in mediating the salinity stress in S. maximus. The expression levels of AMPK α1 and α2 mRNA were a positive correlation with gill Na+, K+-ATPase activities. These findings will aid our understanding of the molecular mechanism of juvenile turbot in response to environmental salinity changes. 相似文献
6.
Pacific abalone Haliotis discus hannai (Haliotidae, Gastropoda) is an economically important shellfish species in northern China. The complete nuclear ribosomal DNA (nrDNA) of Pacific abalone was amplified, sequenced, and analyzed. The length of the nrDNA was determined to be around 10.7 kb, and to contain, in order, small subunit ribosomal RNA (nrSSU) genes (1871 bp), internal transcribed spacer (ITS, 759–762 bp), large subunit ribosomal RNA (nrLSU) gene (3411 bp), and an intergenic spacer (IGS, 4624–4654 bp). The SSU and LSU regions were almost identical in different individuals, and show little variation from those of other abalone species. The two different variations of the ITS2 region were presented, and this phenomenon also existed in other species. A phylogeny tree was constructed, based on ITS region sequence datasets, to determine the evolutionary relationships of abalones. Abalones have two major subclades, mainly distributed in the North Pacific, Europe and Australia. The IGS region of the nrDNA was sequenced and analyzed for the first time. Several repeat fragments were present upstream of the sequence, and were significantly different between individuals (93.86% sequence identity). The complete nrDNA sequence will be useful for the classification, identification, phylogeny, germplasm management, and breeding of this shellfish. 相似文献
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8.
为初步研究中国明对虾MKK4的生物学功能,采用RACE技术克隆获得中国明对虾MKK4基因全长cDNA序列,并对该序列进行分析。结果显示,中国明对虾MKK4基因全长为2 064 bp,开放阅读框长1 221 bp,5'非编码区长214 bp,3'非翻译区长629 bp。将该基因命名为FcMKK4。推测该基因编码406个氨基酸,预测分子量为45.94 ku,理论等电点为8.50。同源性和系统进化分析发现FcMKK4与肩突硬蜱和印度跳蚁的同源性分别为80%和78%,与其他节肢动物MKK4聚为一支。荧光定量RT-PCR结果表明,FcMKK4基因在肌肉中的相对表达量最高,其次为肝胰腺。氨氮胁迫后该基因在中国明对虾肌肉、肝胰腺、血细胞、鳃、心脏、肠和胃中的表达量均显著增加,并有不同的时空表达谱式,表明FcMKK4可能参与中国明对虾非生物胁迫的应答反应。 相似文献
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通过简并引物和RACE技术分离得到了半滑舌鳎Vasa基因的cDNA序列,其全长为2 602 bp,编码包含722个氨基酸残基的蛋白质。半滑舌鳎Vasa在N末端含有5个RGG重复和10个RG重复,具备DEAD-box家族特有的8个保守基序,与东方蓝旗鲔的同源性最高(91%)。利用RT-PCR技术研究了csVasa在性成熟半滑舌鳎中的表达模式。组织分布研究发现,Vasa在半滑舌鳎的卵巢和精巢中表达丰富,心脏中有微量表达;繁殖周期中,卵巢中Vasa的表达量在初级卵黄期和囊泡期要明显高于核周期和退化吸收期;精巢中,Vasa在精子细胞增殖期的表达量要显著高于精子成熟期和精母细胞增殖期。结果表明,Vasa可能在半滑舌鳎的配子发生中起重要作用。 相似文献
11.
瘤背石磺线粒体基因组全序列分析 总被引：1，自引：0，他引：1
采用LA-PCR技术对瘤背石磺线粒体基因组全序列进行了测定和分析。结果表明,瘤背石磺线粒体基因组序列全长13 957 bp,由22个tRNA、2个rRNA、13个蛋白编码基因和19个长度为2~138 bp的非编码区组成。4个蛋白质编码基因和8个tRNA基因从L链编码,其余基因均从H链编码。蛋白质基因的起始密码子,除ND2为TTG以外,均为典型的起始密码子ATN。COⅢ和Cytb基因使用了不完全终止密码子T,其余基因均使用典型的TAA或TAG。预测了22个tRNA基因的二级结构,发现tRNASer缺少DHU臂,tRNASer和tRNAThr的反密码子环上有9个碱基,而不是通常的7个碱基。最长的非编码区含有类似于tRNA的二级结构。基于线粒体基因组编码的13个蛋白质的氨基酸序列,用NJ、MP、ME和UPGMA法构建系统进化树。分析6种软体动物之间的亲缘关系,结果与传统的系统分类基本一致。研究初步确定瘤背石磺与平疣桑椹石磺的亲缘关系比与凯尔特石磺的亲缘关系近。 相似文献
12.
三角帆蚌HcCUBDC基因cDNA的全长克隆与表达分析 总被引：1，自引：1，他引：0
利用cDNA末端快速扩增（Rapid Amplification of cDNA Ends,RACE）方法,获得了三角帆蚌HcCUBDC基因的全长cDNA序列,共5158bp,开放阅读框（ORF）1920bp,编码639个氨基酸,5′端非编码区576bp,3′端非编码区2662bp,GeneBank登陆号为KP067952。生物信息学分析表明,三角帆蚌HcCUBDC蛋白含有一个由19个氨基酸组成的信号肽和4个CUB结构域,但未能比对上已知的蛋白。经荧光定量PCR检测,HcCUBDC基因在紫色和白色蚌前端缘膜、后端缘膜、中央膜、鳃、斧足、肝胰腺、肠和肾8个组织中均有表达,并且都是肝胰腺表达量最低。在紫色蚌中,后端缘膜表达量极显著高于前端缘膜;在白色蚌中,前端和后端缘膜之间表达差异不显著。HcCUBDC基因在紫色蚌后端缘膜中的表达量极显著高于白色蚌,在紫色和白色蚌前端缘膜中的表达量差异不显著。外套膜原位杂交结果显示,HcCUBDC基因主要在缘膜外上皮细胞中表达。研究表明,HcCUBDC基因在三角帆蚌内壳色形成中发挥作用,可为进一步深入研究该基因在珍珠颜色形成过程中的功能及其调控机理提供基础资料。 相似文献
13.
Qian Ma Yong-Quan Su Jun Wang Zhi-Meng Zhuang Qi-Sheng Tang 《Fish physiology and biochemistry》2013,39(2):131-142
Major histocompatibility complex (MHC) plays an important role in the immune response to antigenic peptides in vertebrates. In this study, the full length of MHC IIB cDNA was isolated from the Whitespotted bambooshark (Chiloscyllium plagiosum) by homology cloning, and the rapid amplification of cDNA ends polymerase chain reaction. As a result, the MHC IIB cDNA is 1,407 bp, which contains an open reading frame (ORF) of 831 bp encoding a protein of 276 amino acids. Furthermore, seven alleles of the complete MHC IIB ORF were detected and the variable sites were mainly located in the immunoglobulin-like (β2) region. Tissue distribution analysis showed that MHC IIB can be detected in all the ten tissues examined, with the highest expression in the spleen and gill. Challenge of C. plagiosum with the pathogenic bacteria, Vibrio harveyi, resulted in significant changes in the expression of MHC IIB mRNA in the three immune-related tissues (gill, liver and spleen). These results show that the MHC IIB plays an important role in response to bacterial infection in elasmobranches. 相似文献
14.
Sigang Fan Chaoqun Hu Lvping Zhang Hongyan Sun Jing Wen Peng Luo 《Aquaculture Research》2012,43(9):1306-1316
The complete sequence of mitochondrial DNA (mtDNA) from Stichopus sp. (Echinodermata: Holothuroidea: Stichopodidae: Stichopus) was acquired using conventional PCR and long PCR followed by cloning and sequencing. The mtDNA is a circular molecule of 16 257 bp in length, containing the set of 37 genes, including 13 protein‐coding genes (PCGs), 22 tRNA genes and two ribosomal RNA genes. The plus strand consists of 30.9% A, 23.7% C, 16.0% G and 29.3% T bases (AT skew = 0.027; GC skew = ?0.194). All 13 PCGs encode a total of 3782 amino acids and all 22 tRNA genes were predicted to be capable of folding into a clover‐leaf secondary structure. Intergenetic regions in the mitochondrial genome of Stichopus sp. contain 903 bp in total, with the largest continuous region (674 bp, AT% = 58.6) between tRNA‐Thr and tRNA‐Pro. Analysis of phylogenetic relationship of family Stichopodidae and genetic distances (species among the family Stichopodidae and species within Stichopus monotuberculatus group) based on the partial cox1 sequence demonstrates that Stichopus sp. from the South China Sea is a member of S. monotuberculatus complex. 相似文献
15.
本研究通过高通量测序技术对引入养殖的俄罗斯红参(Apostichopus japonicus)线粒体全基因组进行随机测序并分析,结果显示,俄罗斯红参线粒体基因组全长为16111 bp,共含有37个基因,包括22个tRNA、2个rRNA (l-rRNA和s-rRNA)和13个蛋白编码基因。全序列共存在24处基因间隔和5处基因重叠区。蛋白编码基因与全序列的碱基使用均表现出A、T偏倚。13个蛋白编码基因的密码子完整,除ND1以GTG为起始密码子外,其他均以ATN为起始密码子;终止密码子都是完整密码子,除ND2与ND4的终止密码子分别为TTG与TAG外,其他终止密码子以TAA为主;氨基酸频率最高为丝氨酸Ser(S),其次为亮氨酸Leu(L)。22个tRNA长度范围为66~72 bp,2个基因存在重复,且21个基因能预测其三叶草结构。l-rRNA与s-rRNA的长度与位置不同,对碱基的使用相似。蛋白编码基因的基因排列与海参纲(Holothuroidea)、海胆纲(Echinoidea)排列顺序相同,并与海星纲(Asteroidea)及蛇尾纲(Ophiuroidea)共享基因模块。BLAST分析表明,俄罗斯红参与仿刺参(Apostichopus japonicus)同源性最高。不同海参遗传距离分析发现,俄罗斯红参与3个海域仿刺参最为接近。基于最大似然法分别根据氨基酸与核苷酸序列构建的系统发生树显示,基于核苷酸序列的建树结果更符合亲缘关系较近的海参种类间的分析。引入养殖俄罗斯红参线粒体全序列结构与分析可为后续俄罗斯红参的遗传研究及种质应用提供支持。 相似文献
16.
ABSTRACT: The complete mitochondrial genome sequence of 10 walleye pollocks, Theragra chalcogramma , from the Japan Sea and Bering Sea was determined. The 16 568–16 571 bp genome contains the same 37 mitochondrial structural genes (two ribosomal RNA, 22 transfer RNA, and 13 protein-coding genes) as found in all other vertebrates analyzed, in an organization identical to that of other bony fish. The major non-coding region had several conserved sequence features. Nucleotide variations of ND1, ND5, and control region were high, and these regions appear to be good candidates for high-resolution markers in population studies. 相似文献
17.
大弹涂鱼(Boleophthalmus pectinirostris)是一种生活在潮间带的淤泥滩和红树林等两栖环境中的鱼类,其免疫系统面临比水生生活更大的选择压力。Toll样受体基因(简称TLR)是重要的先天免疫成员,一直是鱼类分子免疫学的研究热点之一。为了探究大弹涂鱼TLR基因是否因为其独特的生活环境而产生适应性进化以及其TLR基因在受到细菌攻击后的免疫应答模式,本研究从大弹涂鱼皮肤转录组中获得了TLR5, TLR8和TLR9完整序列以及TLR3和TLR7部分序列,采用分子生物信息学对大弹涂鱼TLR5, TLR8和TLR9基因序列以及氨基酸序列进行了分析,并根据所构建的系统发育树对5个TLR基因进行了分子进化分析,采用荧光定量PCR方法对大弹涂鱼5个TLR基因的组织表达分布和鳗弧菌攻击后5个TLR基因的免疫应答模式开展了研究。结果显示, TLR5基因全长3071 bp,包括长度为2646 bp的编码区,共编码882个氨基酸;TLR8基因全长3175 bp,包括长度为3033 bp的编码区,共编码1011个氨基酸;TLR9基因全长3398 bp,编码区长度为3093 bp,共编码1031个氨基酸。大弹涂鱼3个TLR基因与其他物种的TLR基因结构相似,具有高度保守性。位点模型结果表明,鱼类TLR3, TLR5和TLR8是高度保守的,而TLR7和TLR9在长期进化过程中产生了适应性进化;而进化枝-位点模型结果表明,为了适应更加复杂多变的两栖环境,大弹涂鱼TLR9基因可能产生了适应性进化。大弹涂鱼5个TLR基因在8个健康组织(肠,眼,肾,肝,脑,肌肉,脾和皮肤)中均有表达,在肝脏和脾脏中的表达量较高。在受到鳗弧菌(Vibrio anguillarum)攻击后的免疫表达模式表明了大弹涂鱼5个TLR基因在应对细菌入侵时起到了重要作用。 相似文献
18.
Chun Xiao Huang Xin Lan Wei Nan Chen Jie Zhang Li Ping Chen Wei Min Wang Jun Yan Li Huan Ling Wang 《Fish physiology and biochemistry》2014,40(1):193-203
Growth differentiation factor 9 (GDF9) is a member of the transforming growth factorβ superfamily and plays an essential role during follicle maturation in mammals. In the present study, the full-length complementary DNA (cDNA) of gdf9 was obtained from Megalobrama amblycephala. The cDNA sequence is 2,061 bp in length with an open reading frame of 1,287 bp encoding 428 amino acid residues. The deduced amino acid sequence shared identities of about 42–86 % with the homologues of other vertebrates. During the early development of embryos, the gdf9 mRNA was detected in zygote with significantly high level and declined sharply by 47 and 87 % at 4 hours post-fertilization (hpf) and 6 hpf and even to an undetectable level through advancing stages. Expression analysis based on quantitative real-time PCR revealed that gdf9 mRNA was mainly expressed in ovary, but much lower levels were also found in some nonovarian tissues. Within the follicle, gdf9 mRNA was localized both in the oocytes and the follicle layer cells by in situ hybridization. During the ovarian cycle, gdf9 mRNA significantly decreased after the previtellogenic stage and became to increase again after the fully grown stage. The results imply that Gdf9 may play critical physiological functions in M. amblycephala early embryonic development and reproduction. 相似文献
19.
Masaru Matsuda Yoshitaka Nagahama Tohru Kobayashi Chika Matsuda Satoshi Hamaguchi Mitsuru Sakaizumi 《Fish physiology and biochemistry》2003,28(1-4):135-139
Although the sex-determining gene Sry has been identified in mammals, no comparable genes have been found in non-mammalian vertebrates. To clone positionally the sex-determining region of the medaka, Oryzias latipes, we generated a Y congenic strain to highlight the genetic differences between the X and Y chromosomes from inbred strains of medaka. We used recombinant breakpoint analysis and deletion analysis of the Y chromosome of a congenic XY female to restrict the sex-determining region to 250-kb stretch of the Y chromosome. Shotgun sequencing of this region predicted 27 genes. Three of these genes were expressed during sexual differentiation. However, only one gene was Y specific. The full-length cDNA sequence of this gene encodes a putative protein of 267 amino acids, including the highly conserved DM domain. We thus named it DMY. To establish a role for DMY during sexual differentiation, we screened wild medaka populations for naturally occurring DMY mutants. Two XY females with distinct mutations in DMY were found in separate populations. The first heritable mutant – a single insertion in exon 3 and the subsequent truncation of DMY – resulted in all XY female offspring. Similarly, the second XY mutant female showed reduced DMY expression with a high proportion of XY female offspring. Furthermore, during normal development, DMY is expressed only in somatic cells of XY gonads. These findings strongly suggest that the sex-specific DMY is required for normal testicular development and is a prime candidate for the medaka sex-determining gene. 相似文献
20.
采用RACE方法,克隆了背角无齿蚌抗菌肽theromacin基因的cDNA全序列。结果显示,该cDNA序列全长为870 bp,5'端非翻译区104 bp,3'端非翻译区460 bp,开放阅读框长为306 bp,共编码101个氨基酸,相对分子量为11 166.9 u。同源性分析显示,该基因编码的蛋白与三角帆蚌theromacin基因氨基酸序列的相似度最高,为73%;与贻贝中发现的抗菌肽defensins、mytilins、myticins和mytimycins等4种类型相似度较低,属于Macin家族(登录号:KJ598604)。实时荧光定量PCR分析结果显示,该基因在血液、肝脏、外套膜、鳃、斧足和肠等组织中均有表达,在外套膜中的表达量最高,在其他组织中的表达量较低;经嗜水气单胞菌诱导后,各个组织的表达量出现明显上调的趋势且与对照组显著差异,推测theromacin基因可能在背角无齿蚌的免疫反应中起重要作用。 相似文献