RESUMO
To identify the genes encoding fatty acid elongases for the biosynthesis of polyunsaturated fatty acids (PUFAs), we isolated a cDNA via degenerate PCR and RACE-PCR from Acanthopagrus schlegelii with a high similarity to the ELOVL5-like elongases of mammals and fishes. This gene is termed AsELOVL5 and encodes a 294 amino acid protein. When AsELOVL5 was expressed in Saccharomyces cerevisiae, it conferred an ability to elongate γ-linolenic acid (18:3 n-6) to di-homo-γ-linolenic acid (20:3 n-6). In addition, the transformed cells converted arachidonic acid (20:4 n-6) and eicosapentaenpic acid (20:5 n-3) to docosatetraenoic acid (22:4 n-6) and docosapentaenoic acid (22:5 n-3), respectively. These results indicate that the AsELOVL5 gene encodes a long-chain fatty acid elongase capable of elongating C(18)Δ6/C(20)Δ5 but not C(22) PUFA substrates.
Assuntos
Ácidos Graxos Dessaturases/genética , Ácidos Graxos Dessaturases/metabolismo , Ácidos Graxos Insaturados/metabolismo , Dourada , Animais , Clonagem Molecular , Expressão Gênica , Dados de Sequência Molecular , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Análise de Sequência de DNA , Ácido gama-Linolênico/metabolismoRESUMO
Delta 6-fatty acid desaturase (D6DES) is used in the synthesis of polyunsaturated fatty acids (PUFAs) from microorganisms to higher animals, including arachidonic acid (ARA) and eicosapentaenoic acid (EPA). A 1,338 bp full-length cDNA encoding D6DES was cloned from Acanthopagrus schlegeli (AsD6DES) through degenerate- and RACE-PCR methods. A recombinant vector expressing AsD6DES (pYES-AsD6DES) was subsequently constructed and transformed into Saccharomyces cerevisiae to test the enzymatic activity of AsD6DES towards the production of n-6 and n-3 fatty acids. The exogenously expressed AsD6DES produced γ-linolenic acid (18:3 n-6) and stearidonic acid (18:4n-3) at 26 and 36% from exogenous linoleic acid (18:2n-6) and α-linolenic acid (18:3n-3), respectively, indicating that it is essentially a delta 6-fatty acid desaturase.
Assuntos
Ácidos Graxos Insaturados/biossíntese , Linoleoil-CoA Desaturase/genética , Linoleoil-CoA Desaturase/metabolismo , Perciformes/genética , Perciformes/metabolismo , Sequência de Aminoácidos , Animais , Sequência de Bases , Biotecnologia , Clonagem Molecular , Primers do DNA/genética , Ácidos Graxos Insaturados/química , Dados de Sequência Molecular , Filogenia , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Homologia de Sequência de AminoácidosRESUMO
Sebastes longispinis is considered to be a valid species despite its incomplete mitochondrial lineage. To obtain basic information of this species, the complete mitochondrial DNA sequence of S. longispinis was determined by next-generation sequencing. The complete sequence is 16,445 bp in length and comprised of 13 protein-coding genes (PCGs), 2 rRNAs, 22 tRNAs and a control region. The complete mitogenome contains 28.0% A, 26.7% T, 17.1% G and 28.3% C nucleotides, with a slight AT bias (54.6%). The start codon of each PCG is ATG, except for COX1 (GTG). The stop codons found in the PCGs are mainly TAA, except for ND1 (TAG), ND3 (TAG), ND4 (AGA), and Cytb (incomplete termination codon, T). The tRNAs have a typical cloverleaf form, except for tRNA(Ser (AGY)), which lacks the DHU arm. The mitogenome of S. longispinis can be used to address the incomplete mitochondrial lineages in allied species of the Scorpaenidae.
Assuntos
Genoma Mitocondrial , Perciformes/genética , Animais , Códon de Iniciação , Códon de Terminação , DNA/química , DNA/isolamento & purificação , DNA/metabolismo , DNA Mitocondrial/química , DNA Mitocondrial/isolamento & purificação , DNA Mitocondrial/metabolismo , Sequenciamento de Nucleotídeos em Larga Escala , Fases de Leitura Aberta/genética , RNA Ribossômico/química , RNA Ribossômico/isolamento & purificação , RNA Ribossômico/metabolismo , RNA de Transferência/química , RNA de Transferência/isolamento & purificação , RNA de Transferência/metabolismo , República da Coreia , Análise de Sequência de DNARESUMO
Hybrids of Sebastes hubbsi, considered a valid species, have been found in the ocean. To examine the basic characteristics of mitochondrial DNA, we analysed the complete mitogenome of S. hubbsis using next-generation sequencing. The complete mitogenome was 16,453 bp in length and comprised 13 protein-coding genes (PCGs), 2 rRNAs, 22 tRNAs and 1 control region. The complete mitochondrial DNA had 27.9% A, 26.7% T, 17.2% G and 28.3% C, showing a slight AT bias (54.5%). The start codon of all PCGs was ATG, except COX1 (GTG). The stop codons in the PCGs were mainly TAA, except ND1 and ND3 (TAG), ND4 (AGA) and Cytb (incomplete termination codon, T). The tRNAs had a typical cloverleaf structure, except tRNA(Ser (AGY)) which lacked the DHU arm. The complete mitogenome of S. hubbsi can be used for hybridization or on-going speciation studies within allied species in the Scorpaenidae.
Assuntos
Genoma Mitocondrial , Perciformes/genética , Animais , Pareamento de Bases/genética , Sequência de Bases , DNA Mitocondrial/genética , RNA Ribossômico/genética , RNA de Transferência/genéticaRESUMO
Due to variations in coloration as well as other morphological features, the identification of Sebastes pachycephalus has been problematic. The complete mitogenome of S. pachycephalus was acquired using next-generation sequencing. The full genome was 16,415 base pairs (bp) in length, assembling two rRNAs, 22 tRNAs, one control region and 13 protein-coding genes (PCGs). The genome constitutes 27.9% A, 26.3% T, 17.2% G and 28.5% C, showing a slight AT bias (54.2%). All PCGs start with an ATG initial codon except COX1, which contains GTG. Most PCG stop codons were TAA, except for ND1 and ND3 (TAG) and Cytb (incomplete termination codon, T). All tRNAs showed the typical cloverleaf structure, except tRNA(Ser(AGY)), which lacks the DHU arm. The complete mitogenome of S. pachycephalus will help further identification and speciation analyses.
Assuntos
Genoma Mitocondrial , Perciformes/genética , Animais , Composição de Bases/genética , Pareamento de Bases/genética , Sequência de Bases , DNA Mitocondrial/genética , RNA de Transferência/genética , República da CoreiaRESUMO
Sebastes taczanowskii is a subarctic species in the north-western Pacific Ocean. To obtain mitochondrial DNA sequences for phylogeny, the complete mitogenome (16 452 bp in length) of S. taczanowskii was constructed using next-generation sequencing. The circular mitogenome contains 13 protein-coding genes (PCGs), 2 rRNAs, 22 tRNAs and the control region, for which gene structure and positions were similar to those of other Scorpaenidae. The complete mitogenome composed of 27.8% A, 26.5% T, 17.3% G and 28.5% C, with a slight AT bias (54.3%). All PCGs use a typical start codon, ATG, except COX1 (GTG). The terminal codon of PCGs was mainly TAR, with the exceptions of ND4 (AGA) and Cytb (incomplete codon, T). Sebastes taczanowskii was clearly divided from other Scorpaenidae in the phylogenetic tree using 2 rRNA and 13 PCGs. The mitogenome of S. taczanowskii can be useful for constructing the molecular phylogenetic tree within Scorpaenidae.
Assuntos
Genoma Mitocondrial , Mitocôndrias/genética , Perciformes/genética , Análise de Sequência de DNA/métodos , Animais , Composição de Bases , Genes de RNAr , Tamanho do Genoma , Sequenciamento de Nucleotídeos em Larga Escala , Filogenia , RNA de Transferência/genética , República da CoreiaRESUMO
The complete mitogenome of the oblong rockfish Sebastes oblongus was constructed using next-generation sequencing. The full genome was 16,396 bp in length, including two rRNA, 22 tRNA, one control region and 13 protein-coding genes (PCGs). The genome consisted of 28.0% A, 26.4% T, 16.9% G and 28.6% C, showing a slight AT bias (54.4%). All PCGs contained an ATG start codon, excluding COX1 that contained a GTG. All PCGs contained the stop codon TAA, excluding ND3 (TAG stop codon) and Cytb (incomplete termination codon, T). All tRNAs contained the typical clover leaf structure, except for two tRNAs, serine (AGY) and threonine, which lacked the DHU arm. The complete mitogenome of S. oblongus will contribute to genetic analysis of the effective population size for aquaculture.
Assuntos
Genoma Mitocondrial , Perciformes/genética , Animais , Pareamento de Bases/genética , Sequência de Bases , Códon/genética , DNA Mitocondrial/genéticaRESUMO
Sebastes owstoni is a popular fishery species in the north-western Pacific Ocean. To obtain basic information on its mitochondrial DNA (mtDNA), the complete mitogenome of S. owstoni was subjected to the next-generation sequencing. The assembled mtDNA was 16,465 bp in length, comprising 13 protein-coding genes (PCGs), 2 rRNAs, 22 tRNAs and a control region. The complete mitogenome comprised of 27.7% A, 26.6% T, 17.3% G and 28.4% C, indicating a slight AT bias (54.3%). The start codon of all PCGs was ATG, with the exception of COX1 (GTG). The stop codons in the PCGs were mainly TAA, with the exception of ND1 and ND3 (TAG), ND4 (AGA) and Cytb (incomplete termination codon, T). All tRNAs had a typical cloverleaf shape, with the exception of tRNASer (AGY), which lacked the DHU arm. The sequenced mitogenome of S. owstoni can be used in genetics-based management initiatives or for constructing the mitochondrial lineages within Scorpaenidae.
Assuntos
Genoma Mitocondrial , Perciformes/genética , Animais , Composição de Bases , Códon de Iniciação , Códon de Terminação , DNA Mitocondrial/química , DNA Mitocondrial/isolamento & purificação , DNA Mitocondrial/metabolismo , Complexo IV da Cadeia de Transporte de Elétrons/química , Complexo IV da Cadeia de Transporte de Elétrons/genética , Proteínas de Peixes/química , Proteínas de Peixes/genética , Sequenciamento de Nucleotídeos em Larga Escala , Fases de Leitura Aberta/genética , Perciformes/classificação , Filogenia , RNA Ribossômico/química , RNA Ribossômico/genética , RNA de Transferência/química , RNA de Transferência/genética , República da Coreia , Análise de Sequência de DNARESUMO
Sebastes is a diverse genus of fish in the family Scorpaenidae. The larvae and adults in this genus have similar morphological characteristics. To obtain basic information for their identification, the mitogenome of S. steindachneri was sequenced using next-generation sequencing. The mitogenome was 16,450 bp in length, and comprised of 13 protein-coding genes (PCGs), 2 rRNAs, 22 tRNAs, and a control region. The mitogenome is 27.4% A, 21.1% T, 17.5% G, and 28.0% C, showing a slight AT bias (54.5%). ATG was the start codon in all of the PCGs, except COX1 (GTG). The termination codon in the PCGs was TAA, except in ND1 and ND3 (TAG), ND4 (AGA), and Cytb (incomplete termination codon, T). All of the tRNAs had a typical cloverleaf shape, except tRNASer(AGY), which lacked the DHU arm. The mitogenome of S. steindachneri should be useful for identifying larvae and constructing a phylogenetic structure for the Scorpaenidae.
Assuntos
Genoma Mitocondrial , Perciformes/genética , Animais , Códon de Iniciação , Códon de Terminação , DNA Mitocondrial/genética , Conformação de Ácido Nucleico , Perciformes/classificação , Filogenia , RNA de Transferência/química , RNA de Transferência/genética , República da Coreia , Análise de Sequência de RNARESUMO
Sebastes vulpes is considered a valid species despite the presence of extensive hybrids. To obtain the basic genetic intraspecific structure, we analyzed the complete mitogenome of S. vulpes using next-generation sequencing. The complete mitogenome was 16,462 bp in length containing 13 protein-coding genes (PCGs), 2 rRNAs, 22 tRNAs and 1 control region. The complete mitogenome comprised 27.7% A, 26.5% T, 17.1% G and 28.6% C, showing a minor AT bias (54.3%). The initial codon in all PCGs was ATG, excluding COX1 (GTG). The majority of stop codons in the PCGs were TAA, excluding ND1 and ND3 (TAG), ND4 (AGA) and Cytb (incomplete termination codon, T). All tRNAs had a typical cloverleaf structure, excluding tRNA(Ser (AGY)), which lacked the DHU arm. The complete mitogenome of S. vulpes can be used to study hybridisation and on-going speciation.
Assuntos
DNA Mitocondrial/genética , Genoma Mitocondrial/genética , Mitocôndrias/genética , Perciformes/genética , Análise de Sequência de DNA/veterinária , Animais , Composição de Bases/genética , Sequência de Bases , Códon de Iniciação/genética , Códon de Terminação/genética , Tamanho do Genoma/genética , Reação em Cadeia da Polimerase/veterinária , RNA Ribossômico/genética , RNA de Transferência/genética , República da CoreiaRESUMO
The complete mitochondrial genome sequence of Sebastes koreanus was determined by next-generation sequencing. The total genome was 16,499 bp in length, comprising two rRNA, 22 tRNA and 13 protein-coding genes (PCGs), as well as the mtDNA control region. The genome constitutes 27.9% A, 26.6% T, 17.1% G and 28.4% C, showing a slight AT bias (54.5%). Of the 13 mitochondrial PCGs, 12 have an ATG start codon while COX1 had a GTG start codon; the incomplete stop codon (T) was present in the ND3, ND4 and Cytb genes. The anti-G bias was considerably lower at the third codon position of PCGs (8.8%). The complete mitogenome sequence of S. koreanus will contribute to molecular phylogenetic and evolutionary analyses of Sebastinae and to the identification of its genetic populations in the Yellow Sea.
Assuntos
Peixes Venenosos/genética , Genoma Mitocondrial , Animais , Sequência de Bases , DNA Mitocondrial/análise , DNA Mitocondrial/genética , Peixes Venenosos/classificação , Dados de Sequência Molecular , Fases de Leitura Aberta/genética , RNA Ribossômico/genética , RNA de Transferência/genética , República da Coreia , Análise de Sequência de DNARESUMO
The complete mitochondrial genome of the dark-banded rockfish Sebastes inermis was determined using next-generation sequencing, including Sanger sequencing. The full genome was 16,504 bp in length, including 2 rRNA, 22 tRNA and 13 protein-coding genes (PCGs), as well as the control region. The genome was 27.8% A, 26.8% T, 17.1% G and 28.3% C, showing a slight AT bias (54.6%). All PCGs started with an ATG initial codon, except for COX1, which started with a GTG. The stop codon of all PCGs was a TAR, except for Cytb, which contained an incomplete termination codon, T. All tRNAs had the typical clover-leaf structure except tRNA(Ser (AGY)), which lacked the DHU arm. The complete mitogenome of S. inermis will aid the identification and genetic population studies on the genus Sebastes.