International Journal of Molecular Zoology 2024, Vol.14, No.1, 44-53 http://animalscipublisher.com/index.php/ijmz 49 3.3 Methods for validation and functional analysis of GWAS results Although GWAS can effectively identify genetic markers associated with traits, the results require further validation and functional analysis to determine the mechanism of action of specific genes. Validation methods usually include: Bioinformatics analysis: Bioinformatics annotation of gene loci identified by GWAS to determine whether they are located near known functional genes or regulatory elements, or whether they are associated with known biological pathways. Gene expression analysis: analyze the expression patterns of candidate genes in different tissues or under different physiological states by techniques such as quantitative PCR or RNA sequencing to verify their relevance to specific behavioral traits (Figure 2). Figure 2 GWAS study identification of genetic markers and their biological significance Functional validation experiments: using gene editing techniques, such as CRISPR-Cas9, to directly manipulate candidate genes and observe changes in behavioral traits in order to confirm the role of these genes in trait expression. Through the application of these methods, researchers are able to not only confirm the accuracy of GWAS results, but also explore the function and mechanism of action of the relevant genes in depth, providing a more scientific and precise basis for the genetic improvement and management of livestock. In conclusion, the application of GWAS technology in the genetic study of livestock behavioral traits provides valuable tools and insights to reveal the genetic complexity and biological nature of traits. With the continuous progress of molecular genetics technology and the continuous optimization of data analysis methods, GWAS will play a greater role in livestock genetics research in the future. 4 Research Progress and Case Studies 4.1 Summary of progress in GWAS studies of behavioral traits in different livestock species With the wide application of genome-wide association analysis (GWAS) technology, genetic studies on behavioral
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