Construction and characterization of a cDNA expression library from the endangered Hu sheep

Hu, Ping-An; Li, X.-C.; Liu, H.-K.; Guan, Weijun; Ma, Yangyang

doi:10.4238/2014.october.31.16

Cited by 5 publications

(3 citation statements)

References 16 publications

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“…Sheep production is affected mostly by inadequate nutrition in rural areas. As a unique Chinese sheep breed [13], the differences in growth rates and reproductivities indicate differences in their nutritional requirements. Whether dietary energy and protein levels have an interactive effect on rumen fermentation and microbial composition in lambs remains unclear in Hu sheep lambs.…”

Section: Introductionmentioning

confidence: 99%

Ruminal Microbiota and Fermentation in Response to Dietary Protein and Energy Levels in Weaned Lambs

Cui

et al. 2020

Animals

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Supplying sufficient nutrients, such as dietary energy and protein, has a great effect on the growth and rumen development of ruminants. This study was conducted to evaluate the effects of dietary energy and protein levels on growth performance, microbial diversity, and structural and physiological properties of the rumen in weaned lambs. A total of 64 two-month-old Hu lambs were randomly allotted to 2 × 2 factorial arrangements with four replicates and with four lambs (half male and half female) in each replicate. The first factor was two levels of dietary metabolizable energy (ME) density (ME = 10.9 MJ/Kg or 8.6 MJ/Kg), and the second factor was two levels of dietary crude protein (CP) content (CP = 15.7% or 11.8%). The trial lasted for 60 days. A low dietary energy level restrained the growth performance of lambs (p < 0.05). The ruminal concentration of acetate and the ratio of acetate to propionate increased but the propionate concentration decreased significantly with the low energy diet. However, the rumen morphology was not affected by the diet energy and protein levels. Moreover, a low energy diet increased ruminal bacterial diversity but reduced the abundance of the phylum Proteobacteria (p < 0.05) and genus Succinivibrionaceae_uncultured (p < 0.05), which was associated with the change in ruminal fermentation phenotypes. By indicator species analysis, we found three indicator OTUs in the high energy group (Succinivibrionaceae_uncultured, Veillonellaceae_unclassified and Veillonellaceae_uncultured (p < 0.01)) and two indicator OTUs in the low energy group (Bacteroidales_norank and Lachnospiraceae_uncultured (p < 0.01)). In conclusion, these findings added new dimensions to our understanding of the diet effect on rumen microbial community and fermentation response, and are of great significance for establishing the optimal nutrient supply strategy for lambs.

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Section: Introductionmentioning

confidence: 99%

Ruminal Microbiota and Fermentation in Response to Dietary Protein and Energy Levels in Weaned Lambs

Cui

et al. 2020

Animals

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“…The Chinese Hu sheep is an important indigenous breed widely raised in the Taihu Lake area of China. This sheep breed is known for its beautiful lambskin, early sexual maturity, and high fecundity (200–250%), and the sheep was listed as one of the 78 nationally protected domestic animals by the Chinese government in 2000 [ 16 , 17 ]. In sheep production, weaning of lambs can be very stressful for both the dam and the offspring.…”

Section: Introductionmentioning

confidence: 99%

iTRAQ-based quantitative proteomic analysis of alterations in the intestine of Hu sheep under weaning stress

et al. 2018

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When Lambs are weaned off ewe’s milk, metabolic, structural, and functional changes often occur in the small intestine. Because information on the effects of weaning stress on the proteome of the intestine is limited, an animal model was established with eight pairs of twin lambs divided into artificially reared and ewe-reared groups, which was followed by proteome analysis using iTRAQ technology. Changes occurred in the morphology of the intestine and 5,338 proteins in three biological replicates with less than a 1.2% false discovery rate were identified and quantified. Among them, a subset of 389 proteins were screened as significantly up- (143) and down-regulated (246) in artificially reared compared with ewe-reared. According to Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis, the differentially expressed proteins that were strongly down-regulated were enriched in immune system processes, biological adhesion, and metabolic processes. The up-regulated proteins were enriched in gene expression, cellular biosynthetic processes, ribosome and RNA binding in response to weaning stress. A series of proteins associated with intestine morphology and immune function were identified, and levels of the mRNAs encoding these proteins were analyzed by real-time quantitative reverse transcription PCR. The results of this study increased our understanding of the response of lambs weaned off ewe’s milk and helped to determine the mechanisms underlying weaning stress.

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“…Primers for L-FABP were designed using Primer Premier 5.0 (Premier Biosoft, Palo Alto, CA, USA) based on the conserved sequences. The forward primer for L-FABP was 5'-CTCGAGGGAGTAGCCTCATTGCCACC-3', while the reverse primer for L-FABP was 5'-GGATCCCTATGAAATGCGGACTTGTCTAAA-3', according to the methods described by Hu et al (2014). The PCR products were cloned into the TA-cloning vector pGEM …”

Section: L-fabp Gene Clonementioning

confidence: 99%

Biological characterization of liver fatty acid binding gene from miniature pig liver cDNA library

Gao

Zhang

et al. 2015

Genet. Mol. Res.

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ABSTRACT. Liver fatty acid binding proteins (L-FABP) are a family of small, highly conserved, cytoplasmic proteins that bind to long-chain fatty acids and other hydrophobic ligands. In this study, a full-length enriched cDNA library was successfully constructed from Wuzhishan miniature pig, and then the L-FABP gene was cloned from this cDNA library and an expression vector (pEGFP-N3-L-FABP) was constructed in vitro. This vector was transfected into hepatocytes to test its function. The results of western blotting analysis demonstrated that the L-FABP gene from our full-length enriched cDNA library regulated downstream genes, including the peroxisome proliferator-activated receptor family in hepatocytes. This study provides a theoretical basis and experimental evidence for the application of L-FABP for the treatment of liver injury.

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Construction and characterization of a cDNA expression library from the endangered Hu sheep

Cited by 5 publications

References 16 publications

Ruminal Microbiota and Fermentation in Response to Dietary Protein and Energy Levels in Weaned Lambs

Ruminal Microbiota and Fermentation in Response to Dietary Protein and Energy Levels in Weaned Lambs

iTRAQ-based quantitative proteomic analysis of alterations in the intestine of Hu sheep under weaning stress

Biological characterization of liver fatty acid binding gene from miniature pig liver cDNA library

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