The Analysis Of Cloning Expression And Genetic Effects Of WFIKKN2 And MSTN Genes In Anguan Hybrid Cattle

Guizhou Red Angus ×guan hybrid cattle,is a hybrid crossbred to Guanling cattle through the hybrid system of(Red Angus♂×Guanling cattle♀)in order to improve the local cattle cattle in Guizhou Province.WFIKKN2 gene(WAP,Follistatin/kazal,Immunoglobulin,Kunitz and Netrin domain containing-2),Also known as the growth differentiation factor-related serum protein 1(Growth and differentiation factor associated serum protein-1,Gasp1),It is a multi-domain protein that regulates the muscle growth inhibitor in animals.Muscle suppressbin MSTN geno The Muscle suppressor MSTN gene(Myostatin,MSTN),also known as growth differentiation Factor 8(Growth differentiation factor 8,GDF8),is a growth factor that plays a wide negative role in regulating muscle growth and development in animals.The WFIKKN2 gene can bind to the MSTN gene prepeptide protein to regulate the activity of the MSTN gene.Two genes haven’t been studied in the Red Angus×guan hybrid cattle,Guizhou.Taking the Red Angus×guan hybrid cattle,Guizhou in this experiment,the advantages of hybrid improvement were explored by comparing the growth performance of Guanling cattle and Red Angus × guan hybrid cattle of different months.Meanwhile,the WFIKKN2 and MSTN gene coding sequence of Red Angus×guan hybrid cattle were cloned and bioinformatics was analyzed.Analysis growth performance determination data with the WFIKKN2,MSTN genotype to identify functional SNPs sites of research value.In addition,the method of real-time fluorescence quantitative PCR of WFIKKN2 and MSTN in adult heart,liver,lung,blood,spleen,kidney,small intestine,fat,lumbar muscle,back muscle,back muscle,shoulder peak and hind muscles,in order to obtain the expression rules of WFIKKN2 and MSTN genes in Red Angus × guan hybrid cattle.The main research results of the test are shown below:(1)The growth performance of 60 Guanling cattle and Anguan hybrid cattle at age of 6,12,18,and 24 months were measured,10 Guanling cattle and 10 Anguan hybrid cattle were slaughtered.The results showed that the growth performance and slaughter performance of Anguan hybrid cattle population at all stages are better than Guanling cattle and the hybridization advantage is obvious.(2)This experiment successfully cloned the complete sequence of the CDS region of Guizhou Anguan hybrid cattle WFIKKN2 and MSTN genes.The length of the CDS region of the WFIKKN2 gene is 1725,which encodes 574 amino acids and the CDS region of the MSTN gene is 1128 bp in length,which encodes 375 amino acids.Four SNPs in the WFIKKN2 gene were found,which are the first exon c.28G>T,the second exon c.956A>G,c.962A>C,and c.1140T>C,all of which cause m RNA secondary level.The structure has changed.Among them,c.956A>G,c.1140T>C are synonymous mutations,c.28G>T,c.962A>C are missense mutations,resulting in glycine(Gly)>tryptophan(Trp)in the encoded amino acid sequence.The change of Leucine(Leu)>Phenylalanine(Phe)leads to changes in the secondary and tertiary structure of the encoded protein,slightly improving hydrophilicity and increasing stability.The CDS region of the MSTN gene is 1128 bp in length and encodes 375 amino acids.Two SNPs are found,c.1077A>C and c.1083C>T are located in the third exon,which are synonymous mutations and c.1077A>C.increased minimum free energy.(3)In this experiment,the SNPs of 109 Anguan crossbred cattle WFIKKN2 and MSTN gene were analyzed for typing and genetic parameter analysis combined with body measurement data for correlation analysis.It found that there were differences in the growth performance of individuals with different genotypes.Among them,c.28G>T locus genotype has significant differences with body weight and chest width;c.962A>C locus genotype has significant differences with body weight,chest width,tube circumference and shoulder circumference;c.1140T>C locus genotype is significantly different from hip-length and tube circumference;c.1077A>C site polymorphism has significant differences in body weight,tube circumference and shoulder circumference;c.1083C>T site has a significant difference in the tube circumference index.(4)This experiment is based on the expression of WFIKKN2 and MSTN genes in adult Anguan hybrid cattle in the heart,liver,spleen,lung,kidney,small intestine,fat,longissimus dorsi,psoas major,hind leg muscle and acromion.The analysis found that the relative expression of WFIKKN2 and MSTN genes in muscle was significantly higher than that in internal organs and the tissue expression trends of the two genes were similar.