Project description:The proteomic profiles of silky fowl egg yolk (SFEY) and Leghorn egg yolk (LEY) were analysed by bottom-up label-free liquid chromatography tandem-mass spectrometry (LC-MS/MS), aiming to provide a theoretical basis for understanding the proteomic and biological differences between the two yolks and further develop the nutritional and biomedical value of silky fowl eggs.
Project description:Cryopreservation causes significant lethal and sub-lethal damage to spermatozoa. In order to improve freezing outcomes, a comprehensive understanding of sub-lethal damage is required. Cryopreservation induced changes to sperm proteins have been investigated in several species, but few have employed currently available state of the art, data independent acquisition mass spectrometry (MS) methods. We used the SWATH LC-MS method to quantitatively profile proteomic changes to ram spermatozoa following exposure to egg yolk and cryopreservation. Egg yolk contributed 15 proteins to spermatozoa, including vitellogenins, apolipoproteins and complement component C3. Cryopreservation significantly altered the abundance of 51 proteins. Overall, 27 proteins increased (e.g. SERPINB1, FER) and 24 proteins decreased (e.g. CCT subunits, CSNK1G2, TOM1L1) in frozen thawed ram spermatozoa, compared to fresh spermatozoa. Chaperones constituted 20% of the proteins lost from spermatozoa following cryopreservation. These alterations may interfere with both normal cellular functioning and the ability of frozen thawed spermatozoa to appropriately respond to stress. This is the first study to apply SWATH mass spectrometry techniques to characterise proteins contributed by egg yolk based freezing media and to profile cryopreservation induced proteomic changes to ram spermatozoa.
Project description:Liver tissues of Guangxi Ma chicken from 32-week old with m performance, 50-week old with high and low laying performance, and 72-week old with high and low laying performance were collected and sequenced in quadruplicate using RNA-seq. The sequences were double-ended sequenced on the DNBSEQ sequencing platform. The sequence reads were quality controlled and then aligned with genomic sequences using HISAT2 program, quantified by featureCounts program, and gene expression levels were verified by qRT-PCR with SYBR Green detection. The results will be helpful to explore the factors that affecting laying performance from the perspective of yolk synthesis and provide a theoretical basis for improving the egg production of Guangxi Ma chicken.
Project description:Purpose: To elucidated key metabolic pathways important for S. Enteritidis survival in egg white. Methods: RNA sample of Mid-log phase bacteria cultures that grown in M9FeS medium were taken as 0h sample, and bacteria incubated with egg white for 6h 12h and 24h were taken as 6,12,24h RNA sample respectively. Three biological replicates were carried out per sample. RNA sample was sequenced with Illumina HiSeq 2000 System. DESeq2 method was used to calculate the differentially expressed genes. qRT–PCR and gene mutation were used to validate RNA-seq result. Results: A total of 12 samples were sequenced. The average output of each sample was 1.33 Gb, and the average comparison rate with the reference genome was 97.51%. M9FeS and egg whites cultivation resulted in 2400 to 2700 expressed genes accounting for 53 to 58% of the whole genome. In this group, 1240 to 1470 genes had significant levels of log2 fold changes ≥1, (Padj <0.05) compared with control. These accounted for 27.3 to 32.2% of the whole genome indicating a large shift in the transcriptional response to egg whites. qRT–PCR result of 21 selected genes highly consistent with the RNAseq results at all three timepoints. 8 out of 12 DEGs mutants showed decreased survival ability in egg white, suggested that the DEGs from the RNAseq results correlated significantly with the egg white resistance phenotype. Conclusions: Using strand-specific RNA sequencing, we found pathways significantly changed in egg white. Genes involved in SOS-dependent and independent DNA damage repair, alkaline pH adaption, osmotic stress adaption, envelope damage repair including maintaining periplasm homeostasis and reinforcing peptidoglycan layer, nitrogen assimilation, Salmonella pathogenicity island 2, iron absorption and biotin synthesis, were significantly up regulated in bacteria surviving in egg whites. Differentially expressed genes involved in energy metabolism, translation, and cell motility, Salmonella pathogenicity island 1 were significantly down-regulated in egg whites.
Project description:Tryptic peptides and N-glycans were spatially mapped and visualised on formalin-fixed paraffin-embedded (FFPE) endometrial cancer tissue microarrays (TMAs) using an ultrafleXtreme MALDI-ToF/ToF MS instrument (Bruker Daltonics). FFPE egg white was placed either side of each TMA and used as an external control to monitor detector performance and sample preparation.
Project description:We used a transcriptomic approach based on the comparison of the expression between the liver of sexually mature hens versus pre-laying pullets to better appreciate which hepatic proteases and antiproteases are specifically expressed in relation to vitellogenesis. Using a 20K chicken oligoarray corresponding to 12 595 different chicken genes, a total of 582 genes were shown to be over-expressed in the liver at sexual maturity of hens (1.2 to 67 fold- difference). Most of the top ten over-expressed genes are known components of egg yolk or of the perivitelline membrane. The combination of different bioinformatic tools reveals 12 proteases and 3 antiproteases amongst the over-expressed genes, including many predicted proteins with yet unknown functions.