Tissue Expression, Function Prediction of ITPR2 Gene and Effects of Its SNPs on Eggshell Quality in Sansui Duck (Anas platyrhyncha domestica)
TAN Guang-Hui, WU Lei, LI Jie-Zhang, QIN Yuan-Yu, ZHANG Yi-Yu*
Key Laboratory of Animal Genetics, Breeding and Reproduction in the Plateau Mountainous Region, Ministry of Education/College of Animal Science, Guizhou University, Guiyang 550025, China
Abstract Inositol-1, 4, 5-triphosphate receptor type Ⅱ (ITPR2) is a key regulatory factor of calcium ion transmembrane transport activity, which affects eggshell quality. In this study, the tissue expression of ITPR2 gene was detected by qRT-PCR, and the SNP sites of protein domain were identified by PCR purification and direct sequencing combined with DNAStar screening. The genetic effect of SNP sites on eggshell quality was analyzed by general linear model. The results showed that ITPR2 gene was expressed in 11 tissues, and the expression of ITPR2 gene was in the order of pancreas>eggshell gland>glandular stomach>kidney>pectoral muscle>spleen>small intestine>lung>liver>heart>muscle>stomach, and the expression of ITPR2 gene in pancreas and eggshell gland was significantly higher than that in other tissues (P<0.01). Three moderate polymorphisms (0.25<PIC<0.50) were detected in exon 16, intron 14 and intron 37 of ITPR2 gene in Sansui ducks (Anas platyrhyncha domestica), among which there was a strong linkage disequilibrium between g.128767757 A>G and g.128659402 G>T, g.128662092 C>T; g.128659402 G>T and g.128767757 A>G deviated from Hardy-Weinberg equilibrium (P<0.05). Correlation analysis showed that the mutation site g.128662092 C>T in exon 16 affected the eggshell quality, and TT genotype significantly increased the eggshell strength (P<0.05); 3 SNPs produced 4 haplotypes and 10 diplotypes, H4 was the dominant haplotype, H2H4 was the dominant diplotype; combination genotype had different effects on eggshell quality, H1H1 and H1H2 significantly increased the eggshell quality strength (P<0.05). The prediction results of ITPR2 protein function showed that the protein encoded by this gene contained 2 647 amino acids, which belonged to non-soluble and unstable protein; there were 6 N-glycosylation sites, and the second and third structures were mainly α-helix and irregular curl. The results of this study suggested that the CT genotype of ITPR2 gene (g.128662092 C>T) in Sansui duck was beneficial to the enhancement of eggshell strength, and the combination of three SNP loci could significantly improve eggshell quality, which could be used as a genetic marker for eggshell quality selection.
TAN Guang-Hui,WU Lei,LI Jie-Zhang, et al. Tissue Expression, Function Prediction of ITPR2 Gene and Effects of Its SNPs on Eggshell Quality in Sansui Duck (Anas platyrhyncha domestica)[J]. 农业生物技术学报, 2020, 28(4): 720-730.
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