microRNAs (miRNAs) are a family of important small non-coding RNA molecules, which participate in the post transcriptional gene regulation. In this review, the numbers and chromosomal distribution of chicken miRNAs, and the regulation and function of chicken miRNAs in immune, embryo development and virus infection were reviewed. Additionally, the applications of chicken miRNAs were also discussed briefly. We hope it can provide references for further study and use of miRNAs in poultry husbandry fields.
Animals ; Chickens ; genetics ; Embryonic Development ; genetics ; Immunity ; genetics ; MicroRNAs ; genetics ; Virus Diseases ; immunology ; veterinary

To obtain chicken CD40L protein, the cDNA was prepared from chicken splenic cells and used as a template to clone and amplify CD40L by PCR. The target gene was cloned into pFastBac vector to construct a pFastBac-chCD40L donor plasmid. Recombinant plasmid was transformed into DH10Bac and recombinant Bacmid-chCD40L was obtained. The Bacmid-chCD40L plasmid was transfected into sf9 insect cells to obtain His-chCD40L protein. In addition, the target gene was cloned into pQM01 vector to construct a pQM01-chCD40L plasmid, recombinant plasmid was transfected into HEK 293T cells to obtain Strep-chCD40L protein. The chCD40L protein was purified by affinity chromatography, and the concentration of purified chCD40L protein was determined to be 0.01 mg/mL. Primary cells were isolated from the bursal tissue of 3-week old SPF chickens, and the chCD40L protein was added to the culture medium to stimulate cells. The chCD40L could bind to CD40 on B cells as examined by Western blotting, indirect immunofluorescence assay and flow cytometry, suggesting that chCD40L protein is biologically active. We successfully obtained chicken CD40L protein of biological activity, which laid the foundation in the in vitro culture of primary B lymphocytes for the isolation and diagnosis of virulent IBDV.
Animals ; Baculoviridae/genetics* ; CD40 Ligand/genetics* ; Chickens ; Cloning, Molecular ; Genetic Vectors/genetics* ; Recombinant Proteins/genetics*

Animals ; Bacterial Proteins ; genetics ; metabolism ; Chickens ; microbiology ; Escherichia coli ; genetics ; metabolism ; Genotype ; beta-Lactamases ; genetics ; metabolism

In order to determine the changes of bacterial community structure and function in the early, middle and late stage of aerobic composting of chicken manure, high-throughput sequencing and bioinformatics methods were used to determine and analyze the 16S rRNA sequence of samples at different stages of composting. Wayne analysis showed that most of the bacterial OTUs in the three composting stages were the same, and only about 10% of the operational taxonomic units (OTUs) showed stage specificity. The diversity indexes including Ace, Chao1 and Simpson showed a trend of increasing at first, followed by decreasing. However, there was no significant difference among different composting stages (P < 0.05). The dominant bacteria groups in three composting stages were analyzed at the phylum and genus levels. The dominant bacteria phyla at three composting stages were the same, but the abundances were different. LEfSe (line discriminant analysis (LDA) effect size) method was used to analyze the bacterial biological markers with statistical differences among three stages of composting. From the phylum to genus level, there were 49 markers with significant differences among different groups. The markers included 12 species, 13 genera, 12 families, 8 orders, 1 boundary, and 1 phylum. The most biomarkers were detected at early stage while the least biomarkers were detected at late stage. The microbial diversity was analyzed at the functional pathway level. The function diversity was the highest in the early stage of composting. Following the composting, the microbial function was enriched relatively while the diversity decreased. This study provides theoretical support and technical guidance for the regulation of livestock manure aerobic composting process.
Animals ; Manure/microbiology* ; Chickens/genetics* ; Composting ; RNA, Ribosomal, 16S/genetics* ; Soil ; Bacteria/genetics*

In order to determine the adjuvant effects of the chicken IL-2 (ChIL-2) on new generation vaccines, ChIL-2 gene was amplified from ConA-stimulated chicken spleen cells by RT-PCR and was directionally inserted into fowlpox virus (FPV) transferring vector p1175 under the control of FPV early/late promoter (PE/L), resulting in recombinant transferring vector p1175IL2. Then the p1175IL2 plasmid was transfected into chicken embryo fibroblasts (CEF) pre-infected with wild type FPV to generate recombinant fowlpox virus expressing ChIL-2 (rFPV-IL2). By selection of blue plaques on the CEF, overlaid with agar containing X-gal, rFPV-IL2 was obtained and purified. The supernatant from CEF monolayer infected with rFPV-IL2 (M.O.I2.0) after 72 hours was detected for the production of ChIL-2 by XTT/PMS colorimetric assay. About 3.6 x 10(5) u/mL of specific ChIL-2 activity was determined. The results show that rFPV-IL2 can express ChIL-2 effectively. rFPV-IL2 provides us with an effective tool for studying avian immunology as well as a potential vaccine-enhancing agent.
Animals ; Chick Embryo ; Chickens ; Fowlpox virus ; genetics ; Interleukin-2 ; genetics ; pharmacology ; Recombinant Proteins ; biosynthesis ; pharmacology

Infectious bursal disease virus (IBDV) is an important member of the Birnaviridae family. IBUV mainly targets the bursa of Fabricius, the central immune organ of chicken, resulting in chicken infectious bursal disease (IBD). IBD represents one of the great challenges for ongoing development of the poultry industry. Reverse genetics for IBDV emerged over twenty years ago. Since then, the technologies behind virus rescue have continually improved leading to a deep understanding of IBDV gene function and tailored vaccine development. Our lab has also been instrumental in the field of IBDV research. Here we review studies on the pathogenic mechanism and the effective prevention and control of IBD.
Animals ; Birnaviridae Infections ; virology ; Chickens ; Infectious bursal disease virus ; genetics ; physiology ; Poultry Products ; virology ; Reverse Genetics

Twenty-four isolates of Newcastle disease virus (NDV) prevailing during 1997 -- 2005 in China were collected. These isolates were purified by CEF plaque assay and replicated in SPF chicken embryos. The hemagglutinin-neuraminidase (HN) genes of these viruses were cloned and sequenced. The HN gene sequences of thirty-six NDV reference strains in GenBank were also used in this study. The amino acid homologing of these viruses were compared and analyzed. The correlations among different fragments of HN gene were also analyzed. The results indicated that the homology of Chinese field NDV strains was 94.4%-99.4%, but 86.9%-89% compared with LaSota and Clone30, 87.9%-89.9% to F48E9, and 87.2%-96.2% to foreign NDV strains. There had the nearest distances among Chinese NDV isolates as compared with that of the LaSota, Clone30 and F48E9 by the phylogenetic tree. However, the distances of seven foreign NDV isolates were very close to Chinese NDV isolates as compared with these of the other foreign NDV isolates. We also found that all the Chinese field isolates were devoid of glycosylation site in position 538 -- 540. There were good correlations between different length amino acid fragments and the genomes of HN, especially the 5'-terminus first 80aa.
Animals ; Chick Embryo ; Chickens ; China ; HN Protein ; genetics ; Newcastle disease virus ; classification ; genetics ; Phylogeny

OBJECTIVETo establish a convenient and accurate method of DNA molecular marker for the identification of corium stomachium galli.

METHODCytb mtDNA sequences of Gallus gallus domestica and three other species of poultry were downloaded from Genbank. Species-specific PCR primers were designed according to the differential DNA fragments of the cytb genes. PCR tests were performed with the DNAs extracted from G. gallus domestica, three other poultry species and domestic mammal animals.

RESULTThe specific primers of G. gallus domestica could only amplify the cytb mtDNA of G. gallus domestica.

CONCLUSIONThe primers are specific to G. gallus domestica mtDNA and can used to discern Corium stomachium from the false medicine.

Black-bone silky fowl, sweet, pungent, and hot-natured, is one of the valuable domesticated birds with special economic value in China's genebank of poultry breed, which has a long history of medicinal and edible uses. It has the effects of tonifying liver and kidney, replenishing Qi and blood, nourishing yin, clearing heat, regulating menstruation, invigorating spleen, and securing essence. Therefore, it has remarkable efficacy of enhancing physical strength, tonifying blood, and treating diabetes and gynecological diseases. Various local black-bone silky fowl breeds have been generated due to the differences in environmental conditions, breed selection, and rearing conditions in different areas of China, which are mainly concentrated in Taihe, Wan'an, and Ji'an in Jiangxi province and Putian, Jinjiang, and Yongchun in Fujian province. The indigenous chicken breeds in China have different body sizes, appearance, coat colors, etc. The complex lineages lead to extremely unstable genetic traits. The diverse breeds similar in appearance result in the confusion in the market of silky fowl breeds. With the rapid development of molecular biological technology, the genetics of black-bone silky fowls has been intensively studied. This article reviews the research progress of the germplasm resources, genetic diversity, and breed identification of black-bone silky fowl in China at the morphology, chromosome, protein, and DNA levels. Further, it introduces the principles, application status, and limitations of DNA markers such as mitochondrial DNA, microsatellite markers, and SNPs. This review provides a theoretical basis for the mining of elite trait genes and the protection and utilization of local black-bone silky fowl germplasm resources in China.
Animals ; Chickens/genetics* ; DNA, Mitochondrial ; Female ; Genetic Variation ; Microsatellite Repeats ; Polymorphism, Single Nucleotide ; Silk/genetics*

To explore the activity of the pmel core promoter of Bashang long-tail chickens, we constructed dual-luciferase expression vectors and transiently transfected into DF1 cells with Lipofectamine 2000. We measured the luciferase activity with the dual-luciferase detection kit. The 1 268 bp fragment in 5-flanking region of the pmel gene in Bashang long-tail chickens was cloned. The region from -1 200 bp to +68 bp included 2 CpG islands and multiple transcription factor binding sites. We constructed 9 expression vectors with different promoter regions and a mutant vector of the core promoter region of the pmel gene of Bashang long-tail chickens. The core promoter region from -840 bp to +68 bp was identified in the pmel gene. The region from -590 to -525 bp negatively regulated the pmel gene during the transcription process. The -840--590 bp and -525--266 bp regions were positive regulatory regions. The polymorphic sites (-456, -435, -410, -374 and -341) had a significant effect on the promoter activity of the pmel gene.
Animals ; Chickens ; genetics ; Cloning, Molecular ; CpG Islands ; Luciferases ; Promoter Regions, Genetic ; gp100 Melanoma Antigen ; genetics