Study On Fatty Acid Difference Between Chahar Sheep And Wuzhumuqin Sheep Based On Fingerprint And Proteomics

Fatty acids in meat are important nutritional and flavor substances,and their differences are influenced by a variety of factors.Differences between breeds are one of the important factors affecting fatty acid content,and there are more and more studies on fatty acid differences between breeds at home and abroad,but relatively little research has been done on sheep.Studies on fatty acid fingerprinting of different breeds of sheep meat and their deposition and metabolism-related protein studies are even less,and there is still a gap in the study of Chahar sheep.In this experiment,we constructed fatty acid fingerprints of the longest dorsal muscle of Ujumqin,Dupo,Chahar and Suffolk sheep,and explored the proteomic factors causing the differences in fingerprint profiles based on similarity differences,for later analysis of the species identification of sheep for meat in Inner Mongolia and the differences in intramuscular fat deposition between Ujumqin and Chahar sheep.In this study,the similarity of traditional Chinese Medicine(TCM)was analyzed by GC-MS chromatographic fingerprints,to established four sheep species fatty acid fingerprints in Inner Mongolia.In view of the differences in fingerprint similarity,the proteome analysis of the differences in fatty acids of Longissimus muscle between Ujimqin sheep and Chahar Sheep was further investigated by using the UP-LC technique.At last,Western Blot technique was used to calculate the difference of protein expression,and the result was compared with SWATH technique.The experimental results were as follows:The conclusion of test 1: After the construction of fingerprint profiles of four meat sheep breeds in Inner Mongolia,it was found that there were 10 common peaks that could be used as discriminatory peaks,and after the similarity test,it was found that Ujumqin and Chahar sheep had the lowest similarity,Dupo and Chahar sheep had the highest similarity.After the results of principal component analysis,it was verified that the method was feasible.The conclusion of test 2: After testing at 1% FDR(false positive),784 total proteins,10897 peptides and 47755 profiles were found in the longest dorsal muscle of Tsakhar sheep.803 total proteins,9850 peptides and 58007 profiles were found in the longest dorsal muscle of Ujumqin sheep.After GO enrichment analysis: Chahar sheep enriched 106 biological functions,74 cytological functions and 40 molecular functions.Ujumqin sheep enriched a total of 94 biological functions,71 cytological functions and 40 molecular functions.After the KEGG analysis : Chahar sheep enriched a total of 81 signaling pathways.Ujumqin sheep enriched a total of 69 signaling pathways The conclusion of test 3: Screening of differential proteins: A total of 182 differential proteins were screened,including 73 up-regulated proteins and 109down-regulated proteins in the longest dorsal muscle of Chahar sheep.GO analysis of differential proteins: BP mainly enriched 45 function;CC Enrichment with 29 component functions;Enrichment of 18 functions on MF.KEGG enrichment analysis:30 pathways were enriched.Only one pathway associated with fatty acid metabolism is included in,3 proteins involved,for ACATI,ECHS1 and HADHB.And all are highly expressed on the Chahar sheep.Through the interaction of protein network,the relationship between the three is found.The conclusion of test 3: The expression of ACAT1 protein in the dorsal longest muscle of Ujumqin sheep and the dorsal longest muscle of Chahar sheep was verified by Western Blot technique,and it was found that ACAT1 protein was highly expressed in the dorsal longest muscle of Chahar sheep,and the test results were consistent with the results of Study 1 and Study 3.The results provide a reference for the later study of intramuscular fatty acid metabolism among different species.In conclusion.with the fingerprint of the longest dorsal muscle of the Ujumqin,Dupo,Chahar and Suffolk sheep could be distinguish.The protein causing the difference in fatty acid fingerprinting between the longest dorsal muscle of Ujumqin and Chahar sheep was then investigated according to the maximum similarity difference.The difference protein is ACATI,ECHS1 and HADHB.It can provide a reference for the identification of sheep breeds for meat in Inner Mongolia at a later stage.