Correlation Analysis between 5'UTR Genetic Variations of Nervous System-specific Transcription Factor Gene NHLH2 and Growth Traits in Jiaxian Red Cattle (Bos taurus)
ZHANG Jing-Min1, WANG Yan-Hong2, SONG Cheng-Chuang2, CHEN Xi2, ZHANG Chun-Lei2, FANG Xing-Tang2,*
1 Shanghai Vocational College of Agriculture and Forestry, Shanghai 201600, China;
2 School of Life Science, Jiangsu Normal University, Xuzhou 221116, China
Abstract：Nescient helix-loop-helix 2 (NHLH2), as nervous system-specific transcription factor, can not only regulate the number of nerve fibers in adipose tissue, but also play an important role in maintaining the energy balance of animals. Searching for DNA markers related to growth traits can provide scientific basis for the protection, development and utilization of beef cattle (Bos taurus) genetic resources. In this study, the genetic variations in 5'UTR of NHLH2 gene and their association with growth traits in Jiaxian red cattle (B. taurus) were explored by the techniques of PCR-single strand conformation polymorphism (PCR-SSCP), DNA sequence analysis and bioinformatics. The 5'UTR of NHLH2 gene was amplified by PCR using the genomic DNA of 374 different individuals of Jiaxian red cattle as templates, and about 400 bp amplified products were obtained. It was found that the 5'UTR of NHLH2 gene was polymorphic by SSCP analysis. Three SSCP genotypes were identified and denominated as genotype AA, AB and BB, respectively. After sequencing the polymorphic products, it was found that there were 2 SNPs (25G>C and 179G>A) in the 5'UTR of NHLH2 gene, which constructed 2 alleles (A and B). A total of 133 amino acids were encoded in bovine NHLH2 gene. The result of homology analysis showed the high similarity of amino acid sequences among different species, which confirmed that the gene was very conservative in the evolution process. The linkage disequilibrium analysis of 2 SNPs showed that the linkage disequilibrium parameter r2 was 1, which indicated that the 2 SNPs were completely linked without cross or recombination. The result showed that there were 2 alleles A and B in the locus with corresponding frequencies of 0.239 and 0.761. And the frequencies of genotype AA, AB and BB were 0.139, 0.201 and 0.660, respectively. The frequency of allele B was higher than that of allele A. Allele B was the dominant allele, and genotype BB was the dominant genotype. The χ2-test value was 0.904, which suggested that the 5'UTR locus of NHLH2 gene was in Hardy-Weinberg equilibrium in this experimental population of Jiaxian red cattle. The polymorphism information content (PIC) value was 0.298, which indicated that the polymorphic locus belonged to moderate polymorphism. Using SPSS 19.0 software and the least square method, the correlation between different genotypes of polymorphic locus and growth traits in Jiaxian red cattle was analyzed. The polymorphisms for 5'UTR locus of NHLH2 gene had significant association (P<0.05) with chest circumference, body length and body weight, while had no significant association with other growth traits. The growth trait indexes of genotype BB individuals were significantly higher than those of genotype AA individuals (P<0.05). The above results confirmed that the correlation was detected between 5'UTR mutation of NHLH2 gene and the growth traits of Jiaxian red cattle, and the polymorphic locus could be used as a molecular marker for the growth traits, which could be used for improvement and breeding of Jiaxian red cattle.
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