Screening of Candidate Proteins Regulating Egg Green-shell Traits Based on TMT Quantitative Proteomics Technology
[Objective]As an important economic trait of green-shell eggs,eggshell color not only affects consumers'purchase preference,but also directly affects the market pricing of green-shell eggs,and the color of green-shell eggs is regulated by multiple genes.The purpose of this study was to explore the candidate proteins and key signaling pathways that regulated the color of green-shell eggs,and provide materials for optimizing the selection and breeding of green-shell chickens and improving the economic value of green-shell eggs.[Method]Chishui Black-bone chickens in the high-yield period of the pure line were used as the research objects,the differentially expressed proteins in the eggshell gland tissues of Chishui Black-bone chickens producing light green eggs(QL)and dark green eggs(SL)were analyzed by tandem mass tags(TMT)quantitative proteomics technology.GO function and KEGG pathway enrichment analysis were performed to screen candidate proteins that regulated the color depth of green shell eggs and verify them by Western blotting.[Result]A total of 5 958 quantifiable proteins were identified,including 102 differentially expressed proteins,of which 46 were up-regulated and 56 were down-regulated.GO functional analysis showed that differentially expressed proteins were mainly enriched in the receptor signaling pathway,vitamin A response and response to estrogen in biological process.Molecular function was primarily enriched in the items of serine hydrolase activity,carbohydrate transmembrane transporter activity,retinol-binding,etc.Cellular component was mainly enriched in items such as egg yolk,collagen trimer and blood particles.Six pathways were significantly enriched in KEGG pathway analysis,among which steroid hormone biosynthesis pathway might change the formation of eggshell color by affecting the eggshell calcification process.Western blotting results showed that the expression of HSD17B7 and TXNRD2 proteins in eggshell glands of Chishui Black-bone chickens in SL group were extremely significantly higher than that in QL group(P＜0.01),and the expression trend was consistent with the proteomics results.[Conclusion]The significant up-regulation of SLC22A15b,SLC35A,SLC2A1,SLC4A4,HSD17B7 and TXNRD2 proteins,and the significant down-regulation of KNG1 protein in differentially expressed proteins might be relate to the formation of eggshell color.The results could provide reference for elucidating the molecular mechanism of pigmentation in green-shell eggs.
万方数据
维普
