SNPs Identification of OCX-32 Gene and Its Correlation with Eggshell Quality in Changshun Blue-eggshell Chicken (Gallus gallus)
WU Lei, LI Jie-Zhang, TAN Guang-Hui, QIN Yuan-Yu, ZHANG Yi-Yu*, LUO Hua- Lun
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：The eggshell matrix protein ovocalyxin-32 gene (OCX-32) is a specific matrix protein that affects eggshell mineralization. The study aim was to investigate the genetic variation effects of eggshell matrix protein OCX-32 gene on eggshell quality. The eggshell quality had been determined by using Changshun Blue eggshell chicken (Gallus gallus). The SNP of OCX-32 gene was screened by sequencing directly of PCR products to analyze the effects on eggshell quality. The predicted results of the three-level structure were consistent with those of the two-level structure, with only slight changes, which were not obvious. Three moderately polymorphic SNPs were detected in the OCX-32 gene of Changshun Blue-eggshell chicken, located at g.22592362 C>T in intron 1 and g.22599114 C>T in exon 6, respectively. Missense mutations (threonine to methionine) and g.22599127 G > A synonymous mutation, both generated 3 genotypes; The distributions of g.22599114 C>T and g.22599127 G> A mutation loci all deviated from Hardy-Weinberg equilibrium (P<0.05). There were no strong linkage disequilibrium among the 3 mutation sites, and 4 haplotypes and 10 diplotype were found. Bioinformatics analysis was found that g.22599114 C >T and g.22599127 G >A mutations could cause the changes of mRNA free energy and protein structure. The CG type free energy had changed from -339.00 kcal/mol to -338.90 kcal/mol. The TA type free energy had changed from -339.00 kcal/mol to -338.40 kcal/mol which led the stability decreased. The protein sequence analysis results had shown that the α helix had changed from 39.27% to 40.00%, the free curl had changed from 41.09% to 40.36% and the β corner and extension had no change. The results of correlation analysis showed that the effects of the 3 SNPs loci on the shell quality of Changshun Blue-eggshell chicken did not reach a significant level (P>0.05); the egg shape index of the diplotype H3H4 individuals was significantly higher than that of the diplotype H2H3 (P<0.05). Diplotype H4H4 (TTCCGG) had the advantage for improving eggshell strength and eggshell thickness and H1H2 (CTCCAG) were the key diplotype to increase the weight of the eggshell, which revealed that the OCX-32 gene mutation could affect the eggshell quality of Changshun Blue-eggshell chicken, and the detected 3 SNPs had the potential to be applied as genetic markers for eggshell quality selection. This study provides data reference and theoretical support for deeper study on the quality of Changshun Blue-eggshell chicken in future.
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