Cloning and expression analysis of hypoxia-related gene HO in cobia

Xie, Ruiqin; Amenyogbe, Eric; Wang, Wei-zheng; Guo, Zhiyong; Chen, Gang; Huang, Jianbo

doi:10.1007/s10499-020-00611-3

Cited by 9 publications

(3 citation statements)

References 25 publications

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“…It has a fast growth rate and inadequate availability in the wild, along with high market values due to its excellent flesh quality. These characteristics have inspired its production in aquaculture all over Asia, especially in China and other parts of the globe as well (Holt et al, 2007;Benetti et al, 2008Benetti et al, , 2010Zhang et al, 2013;Chen et al, 2016;Huang et al, 2020;Xie et al, 2021). Its production reached to approximately 40,000 tons worldwide in 2015 and 59538 tons in 2018 [Food andAgriculture Organization (FAO), 2015, 2020;Wang et al, 2021].…”

Section: Introductionmentioning

confidence: 99%

Probiotic Potential of Indigenous (Bacillus sp. RCS1, Pantoea agglomerans RCS2, and Bacillus cereus strain RCS3) Isolated From Cobia Fish (Rachycentron canadum) and Their Antagonistic Effects on the Growth of Pathogenic Vibrio alginolyticus, Vibrio harveyi, Streptococcus iniae, and Streptococcus agalactiae

et al. 2021

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Large-scale fish farming faces many environmental stresses, which affect their immune systems, growth performance, and physiological homeostasis, resulting in increase in their susceptibility to infections. Some of the most common bacterial infections of cobia fish (Rachycentron canadum) include streptococcosis, vibriosis, furunculosis and mycobacteriosis, and pastelleurosis. Probiotics could be helpful in reducing or limiting the incidence of severe disease infections or outbreaks. Therefore, the present study aimed to isolate the indigenous bacterial species from healthy cobia fish and then selected 3 strains, including Bacillus sp. RCS1 (MW560712), Pantoea agglomerans RCS2 (MW560713), and Bacillus cereus RCS3 (MW560714) from the gut of juvenile’s cobia having advantageous assets or positive characteristics. Their analysis indicated the presence of similar biochemical profiles and all could effectively utilize carbon sources. The biosafety assessment did not show any pathological symptoms after 10 days of injecting the fish with isolated bacteria. The results showed that all the isolated bacteria in the present study had low auto-aggregation capacity within the first 3 h of incubation. The isolated bacteria showed strong tolerance when exposed to a range of pH. Although asymmetrically, a slow rise in the growth of isolated bacteria was observed within the pH range of 1–8 for RC1, 1–7 for RC2, and 1–6 for RC3. The antagonistic effects of isolated bacterial strains on the development of pathogens, including Vibrio alginolyticus, Vibrio harveyi, Streptococcus iniae, and Streptococcus agalactiae, were investigated using Luria-Bertani (LB) agar plates. All the isolated bacteria exhibited inhibitory effects against the pathogens, including V. alginolyticus, V. harveyi, S. iniae, and S. agalactiae. These isolated bacteria were characterized with a wide range of antagonistic activities, non-hemolytic activities, high survivability after heat-treatments and safety confidence, and antibiotic susceptibility. Generally, the characteristics displayed by these strains indicated that they could be used as potential probiotics in the aquaculture industry.

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Section: Introductionmentioning

confidence: 99%

Probiotic Potential of Indigenous (Bacillus sp. RCS1, Pantoea agglomerans RCS2, and Bacillus cereus strain RCS3) Isolated From Cobia Fish (Rachycentron canadum) and Their Antagonistic Effects on the Growth of Pathogenic Vibrio alginolyticus, Vibrio harveyi, Streptococcus iniae, and Streptococcus agalactiae

et al. 2021

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“…Using the method generally followed in our laboratory 88 , 89 , 91 , 93 , 94 , we used the Transgen Biotech TransZol UP Plus RNA kit (Beijing, China) to extract the total RNA from the midgut of juvenile cobias. We then dissolved the RNA in 120 μL of RNase-free water.…”

Section: Methodsmentioning

confidence: 99%

Effects of autochthonous strains mixture on gut microbiota and metabolic profile in cobia (Rachycentron canadum)

Amenyogbe

Luo

et al. 2022

Sci Rep

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The fish immune system is a topic or subject that offers a unique understanding of defensive system evolution in vertebrate heredity. While gut microbiota plays several roles in fish: well-being, promoting health and growth, resistance to bacterial invasion, regulation of energy absorption, and lipid metabolism. However, studies on fish gut microbiota face practical challenges due to the large number of fish varieties, fluctuating environmental conditions, and differences in feeding habits. This study was carried out to evaluate the impacts of supplemented three autochthonous strains, Bacillus sp. RCS1, Pantoeaagglomerans RCS2, and Bacilluscereus RCS3 mixture diet on cobia fish (Rachycentroncanadum). Also, chromatography, mass spectrometry and high throughput sequencing were combined to explore composition and metabolite profile of gut microbiota in juvenile cobia fed with supplemented diet. In the trial group, juvenile cobia received diets supplemented with 1 × 1012 CFU mL−1 autochthonous strains for ten weeks and a control diet without supplementation. Juvenile cobia receiving diets supplementation exhibited significantly improved growth than those without additives (control). Haematological indices, such as red blood cells, white blood cells, corpuscular haemoglobin concentration, mean corpuscular volume, haemoglobin, and mean corpuscular haemoglobin, were higher in the supplemented group. Similarly, digestive enzymes (trypsin, lipase, amylase, pepsin and cellulose, activities) activities were higher in supplemented diet with an indigenous isolates mixture. Serum biochemical parameters albumin, globulin, and total protein were significantly higher, while triglyceride, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and cholesterol showed no significant difference. On the other hand, glucose was significantly (P < 0.05) higher in the group without supplementation. On gene expression in the midgut, Immunoglobulin, Colony-stimulating factor receptor 1, major histocompatibility complex 1 were up-regulated by native isolates while T cell receptor beta, and Major histocompatibility complex 2 showed no significant difference. Gut bacterial composition was altered in fish receiving supplemented diet with autochthonous strains. Metabolomics also revealed that some metabolic pathways were considerably enriched in fish fed with supplemented diet; pathway analysis based on Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment revealed that differentially expressed metabolites were involved in galactose metabolism, tryptophan metabolism, carbohydrate digestion and absorption, purine metabolism, and ABC transporters. Functional analysis of bacterial community showed that differences in enriched metabolic pathways generally comprised carbohydrate and its metabolites, nucleotide and its metabolites, amino acid and its metabolites, heterocyclic compounds, and tryptamines, cholines, pigments. The current investigation results showed that autochthonous strains mixture has significantly enhanced the growth, survival, and innate and adaptive immunities of juvenile cobia.

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“…However, in most cases, the gene of eukaryote cannot be directly obtained by this method because of containing introns. Although whole-gene synthesis [6] and reverse transcription methods [7] can solve this problem, they are time-consuming, expensive and complicated to operate. Moreover, big genes are more problematic and far more expensive.…”

Section: Benchmarkmentioning

confidence: 99%

Rapid Golden Gate assembly of exons from genomic DNA for protein expression in Escherichia coli and Pichia Pastoris

Cheng

Zhong

et al. 2021

BioTechniques

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The development of a quick, single-step cloning system for generation of multiexon gene expression constructs is presented. The system allows efficient and cost-effective assembly of multiple exons of interest genes into different expression plasmids in both Escherichia coli and Pichia pastoris. The high cloning efficiency and low cost of the system make it ideal for a novel workflow for the assembly of intron-bearing genes for expression in two different expression hosts.

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Cloning and expression analysis of hypoxia-related gene HO in cobia

Cited by 9 publications

References 25 publications

Effects of autochthonous strains mixture on gut microbiota and metabolic profile in cobia (Rachycentron canadum)

Rapid Golden Gate assembly of exons from genomic DNA for protein expression in Escherichia coli and Pichia Pastoris

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