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Xinhong Guo Jinpeng Yan Shaojun Liu Bing Xiang Yun Liu 《Fish physiology and biochemistry》2010,36(2):125-133
To investigate the evolutional significance of Sox9 in fish, we isolated and characterized Sox9a cDNA and genomic clones in triploid crucian carp. The cDNA encoded a protein of 457 amino acids with an HMG box and showed
more than 60% amino acid sequence identity with known vertebrate Sox9 proteins. Triploid crucian carp and vertebrate Sox9s showed similar gene structure, and two introns in the coding region were located at conserved positions. On the basis of
the amino acid sequences, Sox9a can be categorized into the same subgroup of Sox-E proteins as Sox8, 9, and 10. Interestingly, the expression of triploid crucian carp Sox9a was predominantly observed not in the ovary but in the testis by Northern blot and RT-PCR analysis. The expression analysis
of Sox9a suggested that it may seldom contribute to the formation of normal functions of spermatozoa, but it may play an important
role in the development of testicular tubules. Besides the testicular expression, Sox9a was also shown to be expressed in many other tissues including the brain, kidney, and heart of triploid crucian carp, indicating
that Sox9 may have unique functions in some specific tissues during development. 相似文献
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The growth hormone (GH) gene isolated and cloned from various Labeo species (L. rohita, L. calbasu, L. fimbriatus, L. gonius, L. bata, and L. kontius) is shown to contain a single copy in the haploid genome, with an overall size of ∼2.5 kb. The GH gene in all the Labeo species studied has five exons and four introns of various sizes with the exon/intron boundary sequence of GT/AG. The length
variation of the GH gene between the species is found to be due to length variation in the form of several deletions in the
third intron. The length of individual exons is the same in all the species with an open reading frame (ORF) of 630 bp (210
amino acids) except in L. rohita, which has a 9 bp deletion in the fourth exon, resulting in a shorter GH of 621 bp (207 amino acids). The similarity in the
nucleotide and amino acid sequences between the different Labeo species is greater than 97%, in spite of eight amino acids being altered in the GH protein of Labeo that reside outside the conserved domain sequence required for its function. Nucleotide substitutions are seen in the form
of 20 transitions and three transversions in the ORF of the GH gene. Both types of transitions (A–G; T–C) and only one type
of transversion (A–C) are detected in the GH gene. Codon preference in GH gene shows a strong preference for G and C in the
wobble position of the codons. Genetic interrelationships determined between Labeo and other species of fishes using nucleotide sequence of GH cDNA supports the overall teleost classification of Nelson (Fishes
of the World. Wiley, New York, 1984) with separate clades for Ostariophysi, Protacanthopterygii, and Acanthopterygii. Besides, the unweighted pair group method
with arithmetic means (UPGMA) analysis clearly distinguishes between the species having five exons and four introns in the
GH gene from the species having six exons and five introns in the same gene. The Labeo species analyzed in the present study could be clustered into two groups using the maximum-parsimony method on the intron
sequences data of the GH gene. 相似文献
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三疣梭子蟹Sox基因HMG盒的克隆分析 总被引：1，自引：0，他引：1
利用能扩增人类SRY基因HMG盒的一对简并引物,分别对三疣梭子蟹雌雄个体基因组DNA进行扩增,均得到216 bp和约400 bp的两条带,不存在性别差异,通过PCR-SSCP分析,未发现雌蟹或雄蟹所特有的Sox基因。对216 bp的带进行克隆测序得到6个Sox基因,分别命名为PTSox21、PTSox12、PTSox11a、PTSox11b、PTSox11c和PTSox4。其中,PTSox21、PTSox12和PTSox4氨基酸序列与人类Sox21、Sox12和Sox4基因的同源性分别为90%、66%和63%,PTSox11a、PTSox11b和PTSox11c氨基酸序列均与人类Sox11基因有63%的同源性。此外,PTSox11a和PTSox11b的氨基酸序列之间的同源性达到了98.6%,只在第44位氨基酸残基不同。与其他物种Sox基因氨基酸序列的同源性比较发现,PTSox21与饰纹姬蛙Sox2a有92%的同源性,而PTSox12、PTSox11a、PTSox11b和PTSox11c均与意大利蜜蜂的Sox1有73%的同源性,PTSox4与意大利蜜蜂的Sox1有72%的同源性。 相似文献
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Wei Liu Huayu Song Aoyun Li Xinxin Du Yuezhong Liu Yan He Quanqi Zhang Jie Qi 《Fish physiology and biochemistry》2016,42(5):1275-1285
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Two isoforms of the full-length cDNA of the growth hormone receptor (GHR) of the Atlantic salmon (Salmo salar; ss) were cloned by a PCR approach using RACE. Respectively, the cDNA sequences of ssGHR isoforms 1 and 2 are 2654 and 2608 nucleotides long, with 1782 and 1773 nucleotide ORFs. The resulting coded proteins
are 594 and 590 aa long, with 19 and 20 aa signal peptides. The two isoforms share 86% protein and 87% cDNA sequence similarity.
Isoform 1 is most similar to other salmonid GHR isoforms 1 while isoform 2 is most similar to salmonid GHR isoforms 2 (93–95%).
Similarity with other teleost species was lower (37–44%). The bioactivity of the cloned ssGHR was tested by transfecting the ssGHR isoform 1 cDNA into CHO-K1 hamster cells, incubating with recombinant salmon GH (sGH) or native ovine prolactin (oPRL),
and measuring cell proliferation by the MTT assay. The ssGHR-transfected cells significantly increased proliferation when stimulated by sGH at all concentrations. oPRL stimulated
ssGHR-transfected cells at higher concentrations due to receptor cross reaction. ssGHR isoforms 1 and 2 contain a single transmembrane domain and the typical conserved motifs found in other teleost GHRs, including
four paired cysteine residues and five potential N-glycosylation sites in the extracellular domain, Box I and Box II, as well as seven potential tyrosine phosphorylation sites
in the intracellular domain. However, in salmonids, these motifs differ from those of other teleosts, and could be responsible
for differentiated hormone binding, signal transduction and response. 相似文献
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Sex steroid level and sexual dimorphism expression of genes in gonads of the great sturgeon Huso huso Linneaus, 1758 during maturity developmental stages 
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下载免费PDF全文 Mahtab Yarmohammadi Mohammad Pourkazemi Rezvanollah Kazemi Mohammad Ali Yazdani Sadati Ali Hallajian Mohammad Hassanzadeh Saber 《Aquaculture Research》2017,48(4):1413-1429
Little is known about molecular mechanisms involved in gonad differentiation in sturgeon species. In non‐mammalian vertebrates, sox9, dmrt1, cyp17a1 and ar are male specific genes in testicular differentiation and are highly conserved. In order to understand the mechanism underlying testicular development in sturgeon, we investigated sex steroids level of 11‐ketotestosterone (KT) and testosterone (T) and relative expression of sox9, dmrt1, cyp17a1 and ar in great sturgeon gonads during different stages of sexual maturity. The results showed all studied genes had dimorphic patterns in males. In immature gonads (stage I), only cyp17a1 expressed in male gonads, while other genes did not express, and KT and T levels were low. Dmrt1 showed dimorphic expression pattern in male gonads at maturity stage II, III and IV. Furthermore, sox9 and ar mRNA presented significant dimorphic expression pattern in male gonads only at maturity stage 4. Plasma androgens levels were significantly higher in males compared to females during maturity stages II, III and IV. The results showed that among these four genes, only cyp17a1 expressed in male gonads at maturity stage I (immature), suggesting that this gene may be applicable as a sex marker in recently differentiated male great sturgeon. Sexually dimorphic patterns in other studied genes (sox9, dmrt1 and ar) suggesting that these genes may be important for testicular development and differentiation in premature great sturgeon. The obtained results provide a foundation for further research on sex differentiation and developing strategies for the sexing of sturgeon for aquaculture. 相似文献
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采用逆转录聚合酶链反应(RT-PCR)及cDNA末端快速扩增(RACE)技术克隆了中国大鲵(Andrias davidianus)Sox19基因全长cDNA序列,并进行各组织间的表达分析。中国大鲵Sox19基因全长1290 bp,ORF长858 bp,编码285个氨基酸,位于39~109位的HMG-box是其主要功能区。氨基酸序列分析表明,其与舌齿鲈(Dicentrarchus labrax)、斑马鱼(Danio rerio)及红鳍东方鲀(Takifugu rubripes)的相似度分别为64%、58%、55%。采用邻接法对不同物种的Sox19编码区序列构建分子系统树发现:中国大鲵Sox19基因属于较早从鱼类分化出来的基因,表明Sox19基因从中国大鲵到鱼类的进化速度比较快。荧光定量PCR结果显示,Sox19在所有检测的组织中均有表达,且在心脏中的表达量最高,卵巢中次之,肾脏中的表达量略高于肠道。中国大鲵Sox19基因的发现打破了Sox19基因一直被认为是鱼类所特有基因的观点,填补了两栖类有关此基因的空白,为更进一步的了解Sox19基因在中国大鲵中的重要功能奠定了基础,同时Sox19基因还可作为研究中国大鲵早期胚胎神经系统发育情况的分子标记,为其他两栖类动物的研究提供借鉴。 相似文献
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Twist2 在调控动物发育过程发挥了重要作用。为解析鳜(Siniperca chuatsi) twist2 基因的表达模式以及参与肌肉发育调节的功能和机制, 本研究从鳜基因组数据库获得 twist2 基因序列, 采用生物信息学方法对 Twist2 蛋白特征和同源进化进行了分析。鳜 twist2 基因所编码的蛋白由 162 个氨基酸组成, 具有一个保守功能域 bHLH 结构域。 基于氨基酸序列的多序列比对结果显示, Twist2 蛋白在脊椎动物中具有较高的相似性; 采用 RT-qPCR 技术分析了鳜 twist2 基因的时空表达规律, 发现 twist2 基因在不同发育阶段存在表达差异, 原肠期之前表达水平较低, 从神经胚期开始显著上升, 尾芽期表达最高; 在鳜组织中的表达情况显示, 该基因在脾脏中表达量较高, 其次是脑和红肌。采用整胚原位杂交技术检测其早期胚胎不同发育阶段的 twist2 基因定位, 发现其主要在神经胚层和体节中特异性表达。采用 Targetscan 和 RNAhybrid 工具对可能靶向鳜 twist2 基因的 miRNAs 进行预测, 结果发现 miR-30a、 miR-30b、miR-30e-5p 和 miR-204 可能作用于 twist2 3ʹUTR, 提示 twist2 是 miR-30a、miR-30b、miR-30e-5p 和 miR-204 的潜在靶基因。鳜短期饥饿胁迫实验显示, 在饥饿 3 d 时 twist2 的表达与上述 miRNAs 的表达呈明显的负相关, 表明 twist2 与上述 4 个 miRNAs 存在潜在的调控关系。因此, 本研究结果将有助于从分子水平了解 twist2 基因的序列特征、时空表达规律以及其调控肌肉生长发育的潜在功能, 为鱼类发育生物学以及健康养殖提供理论依据。 相似文献
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A homologue of dermatopontin from Haliotis diversicolor and its response to pathogenic infection 下载免费PDF全文
下载免费PDF全文 Guodong Wang Ziping Zhang Shi Lin Lili Zhang Baozhen Wang Shuhong Wang Yilei Wang 《Aquaculture Research》2015,46(7):1537-1549
Dermatopontin (DPT), a component of the extracellular matrix, plays important roles in cell‐matrix interactions and matrix assembly. Some studies have revealed that it has more general functions in biological activities. However, its function in molluscs is poorly understood. In this study, a molluscan DPT gene, saDPT2, was cloned from small abalone Haliotis diversicolor. The full‐length cDNA of saDPT2 sequence is 620 bp, with a 531 bp open reading frame encoding a protein of 177 amino acids (aa). Amino acid sequence analysis revealed that saDPT2 shares conserved signature motifs with other DPT proteins, including three repeats of 10‐residue motif (S‐X‐H‐X‐N‐X‐Y‐E‐D‐R), which is similar to mammalian 6‐residue repeating sequence of D‐R‐E/Q‐W‐X‐F/Y. Quantitative real‐time PCR was employed to investigate the tissue distribution of saDPT2 mRNA, its expression at different developmental stages, and in abalone under bacteria challenge. The saDPT2 mRNA could be detected in all examined tissues and developmental stages. Moreover, the saDPT2 mRNA was up‐regulated in haemocytes and gills after bacteria injection. The results indicate that the saDPT2 could respond to pathogenic infection and may play a role in adult abalone immune system. 相似文献
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M. T. Thébault A. Tanguy A. L. Meistertzheim J. P. Raffin 《Fish physiology and biochemistry》2010,36(4):819-825
AMP-deaminase (AMPD, EC 3.5.4.6), which catalyzes the irreversible hydrolytic deamination of AMP to IMP and ammonia, is an
important energy-related enzyme. The partial genomic sequence of the gene encoding myoadenylate deaminase (AMPD1) from the
teleost fish Platichthys flesus was determined. The amino acid sequence of P. flesus AMPD1 shows 82% homology with that of the teleost fish Danio rerio. Comparison of genomic sequences of P. flesus and Rattus norvegicus reveals a high degree of conservation of both sequence and structural organization. A phylogenetic analysis of AMPD sequences
shows that bony fish and mammalian AMPD1s arise by duplication of a common primordial gene. 相似文献
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Gad Degani 《Aquaculture Research》2004,35(9):807-815
The DNA of two laboratory strains of guppy (Poecilia reticulata), Black Yellow (BYS) and Red Flame (RFS), was studied with respect to their colour differences, in two generations (F1 and F2). Their varying morphological colours were related to their cloned fragments of cytochrome b mitochondrial DNA (mtDNA) and random‐amplified polymorphic DNA‐polymerase chain reaction (RAPD‐PCR). The F1 generation was characterized by various forms. The male BYS could be divided into 68% having black–yellow tails and 14% having black–yellow–red tails. The other variations were found in very low percentages. The percentage of black–yellow‐tailed males increased to 85 in the F2 generation, and the percentages of the various other forms consequently decreased. Only 63% of BYS males had black–yellow dorsal fins in the F1 generation, but this percentage increased to 84 in F2. Compared with the males, fewer variations were found in female colour patterns in the F1 generation. A high percentage of BYS females (81%) colour was found with no significant increase in the F2 generation. However, variations decreased in the F2 females. On the other hand, a very high variation was found in female fins in the F1 generation: only 32% were of BYS colour and 25% had no‐colour fins. However, a significant increase in BYS colour was found in the F2 generation (61%) and 39% had no colour. The variation in RFS was lower than BYS in the F1 generation: 81% of the F1 males had red–yellow tails with colour, 46% of the fins were yellow and 36% were red–white. In females, a very high percentage (84%) had red–yellow tails and 76% had no‐colour dorsal fins. Mitochondria DNA markers and genomic DNA were studied in various laboratory strains. In the clone of the fragments of cytochrome b, the bands correlated to the colour phenotype. A fragment of the cDNA sequence was determined from a 268‐bp cloned with fragments of the guppy cytochrome b mtDNA gene. The genes varied for the two strains in only two base pairs, starting at the nucleotide position 171 and ending at position 174. Three primers showed good results in the RAPD‐PCR and were found suitable for the study of DNA variations in guppy. The high variation detected in BYS, in comparison with RFS, was reflected by changes in band‐sharing (BS) values ranging from 0.66 to 1, versus 0.8 to 1 in RFS. 相似文献