神经系统特异性转录因子基因<em>NHLH2</em>的5’UTR遗传变异与郏县红牛生长性状的相关分析

doi:10.3969/j.issn.1674-7968.2020.08.008

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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.

Key words： NHLH2 (nescient helix-loop-helix 2) 5'UTR Genetic variation Jiaxian red cattle Growth trait

Received: 10 January 2020

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Corresponding Authors: *xtfang@163.com

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	Articles by authors
	ZHANG Jing-Min
	WANG Yan-Hong
	SONG Cheng-Chuang
	CHEN Xi
	ZHANG Chun-Lei
	FANG Xing-Tang

Cite this article:

ZHANG Jing-Min,WANG Yan-Hong,SONG Cheng-Chuang, et al. Correlation Analysis between 5'UTR Genetic Variations of Nervous System-specific Transcription Factor Gene NHLH2 and Growth Traits in Jiaxian Red Cattle (Bos taurus)[J]. 农业生物技术学报, 2020, 28(8): 1410-1416.

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http://journal05.magtech.org.cn/Jwk_ny/EN/10.3969/j.issn.1674-7968.2020.08.008 OR http://journal05.magtech.org.cn/Jwk_ny/EN/Y2020/V28/I8/1410

[1] 陈宏,张春雷.2008.中国肉牛分子育种研究进展[J].中国牛业科学,34(4):1-7.
(Chen H,Zhang C L.2008.Recent progress in beef cattle molecular breeding in China[J].China Cattle Science,34(4):1-7.)
[2] 胡娟,彭小忠.2007.神经系统特异性转录因子Nhlh2的研究进展[J].中国医学科学院学报,29(3) :436-440.
(Hu J,Peng X Z.2007.Research advances in nervous system-specific transcription factor,Nhlh2[J].Acta Academiae Medicinae Sinicae,29(3):436-440.)
[3] 李枫梅.2017.肉用牛bHLH转录因子家族成员的鉴定与分析[D].硕士学位论文,阜阳师范学院,导师:刘武艺,pp.16-23.
(Li F M.2017.Identification and functional analysis of the basic helix-loop-helix (bHLH) transcription factors in the meat cattle[D].Thesis for M.S.,Fuyang Normal University,Supervisor:Liu W Y,pp.16-23.)
[4] 罗莉,魏曙光,常辽,等.2019.乳腺癌BRCA1相互作用蛋白1基因功能区单核苷酸多态性与习惯性流产的相关性[J].西安交通大学学报(医学版),40(6):959-961.
(Luo L,Wei S G,Chang L,et al.2019.Association between BRCA1-interacting protein-terminal helicase 1 gene functional single nucleotide polymorphisms and recurrent miscarriage[J].Journal of Xian Jiaotong University Medical Sciences,40(6):959-961.)
[5] 祁琪,邵文涵,杨灵芝,等.2018.秦川牛mtDNA 12S rRNA和16S rRNA基因多态性及其与生长性状的相关分析[J].中国兽医学报,38(11):2205-2210.
(Qi Q,Shao W H,Yang L Z,et al.2018.Polymorphism of mtDNA 12S rRNA gene and mtDNA 16S rRNA gene and analysis on relationship with growth traits in Qinchuan cattle[J].Chinese Journal of Veterinary Science,38(11):2205-2210.)
[6] 王勇,姚勤,陈克平.2010.动物bHLH转录因子家族成员及其功能[J].遗传,32(4):307-330.
(Wang Y,Yao Q,Chen K P.2010.Progress of studies on family members and functions of animal bHLH transcription factors[J].Hereditas,32(4):307-330.)
[7] 温丽娟,刘发伟,林长光,等.2018.猪Nhlh2基因启动子活性及其表达调控的初步研究[J].畜牧兽医学报,49(4):667-674.
(Wen L J,Liu F W,Lin C G,et al.2018.Analysis of the promoter activity and expression regulation of porcine Nhlh2 gene[J].Chinese Journal of Animal and Veterinary Sciences,49(4):667-674.)
[8] 张婧敏,房兴堂.2015.AQP7基因内含子4多态性与郏县红牛生长性状的关联分析[J].中国畜牧兽医,42(8):2137-2143.
(Zhang J M,Fang X T.2015.Association analysis on polymorphisms of AQP7 gene intron 4 with growth traits in Jiaxian cattle[J].China Animal Husbandry & Veterinary Medicine,42(8):2137-2143.)
[9] 赵芳芳,张鸣明,罗玉柱,等.2019.DHPLC和PCR-SSCP两种方法筛查基因变异的比较[J].中国草食动物科学,39(4):11-14.
(Zhao F F,Zhang M M,Luo Y Z,et al.2019.Comparison of DHPLC and PCR-SSCP in screening gene variation[J].China Herbivore Science,39(4):11-14.)
[10] Al Rayyan N,Wankhade U D,Bush K,et al.2013.Two single nucleotide polymorphisms in the human nescient helix-loop-helix 2 (NHLH2) gene reduce mRNA stability and DNA binding[J].Gene,512(1):134-142.
[11] Al Rayyan N,Zhang J H,Burnside A S,et al.2014.Leptin signaling regulates hypothalamic expression of nescient helix-loop-helix 2 (Nhlh2) through signal transducer and activator 3 (Stat3)[J].Molecular and Cellular Endocrinology,384(1-2):134-142.
[12] Botstein D,White R L,Skolnick M,et al.1980.Construction of a genetic linkage map in man using restriction fragment length polymorphisms[J].The American Journal of Human Genetics,32(3):314-331.
[13] Brennan K M,Vella K R,Good D J.2006.Genetic analysis of NHLH2 and its putative role in bovine body weight control[J].Animal Genetics,37(1):24-27.
[14] Cabanelas A,Lisboa P C,Moura E G,et al.2006.Leptin acute modulation of the 5'-deiodinase activities in hypothalamus,pituitary and brown adipose tissue of fed rats[J].Hormone and Metabolic Research,38(8):481-485.
[15] Coyle C A,Jing E,Hosmer T,et al.2002.Reduced voluntary activity precede adult-onset obesity in Nhlh2 knockout mice[J].Physiology & Behavior,77(2-3):387-402.
[16] Fox D L,Vella K R,Good D J.2007.Energy balance pathways converging on the Nhlh2 transcription factor[J].Frontiers in Bioscience,12(8-12):3983-3993.
[17] Garcia M F,Moore C D,Schulz K N,et al.2019.Structural features of transcription factors associating with nucleosome binding[J].Molecular Cell,75(5):921-932.
[18] Good D J,Braun T.2013.NHLH2:At the intersection of obesity and fertility[J].Trends in Endocrinology and Metabolism,24(8):385-390.
[19] Good D J,Coyle C A,Fox D L.2008.Nhlh2:A basic helix-loop-helix transcription factor controlling physical activity[J].Exercise and Sport Sciences Reviews,36(4):187-192.
[20] Graziano C,Carone S,Panza E,et al.2009.Association of hereditary thrombocythemia and distal limb defects with a thrombopoietin gene mutation[J].Blood,114(8):1655-1657.
[21] Kimchi-Sarfaty C,Oh J M,Kim I W,et al.2007.A 'silent' polymorphism in the MDR1 gene changes substrate specificity[J].Science,315(5811):525-528.
[22] Li W M,Guo B L,Tao K,et al.2019.Inhibition of SIRT1 in hippocampal CA1 ameliorates PTSD-like behaviors in mice by protections of neuronal plasticity and serotonin homeostasis via NHLH2/MAO-A pathway[J].Biochemical and Biophysical Research Communications,518(2):344-350.
[23] Nackley A G,Shabalina S A,Tchivileva I E,et al.2006.Human catechol-O-methyltransferase haplotypes modulate protein expression by altering mRNA secondary structure[J].Science,314(5807):1930-1933.
[24] Nei M.1978.Estimation of average hetorozygosity and genetic distance from a small number of individuals[J].Genetics,89(3):583-590.
[25] Shi Y Y,He L.2005.SHEsis,a powerful software platform for analyses of linkage disequilibrium,haplotype construction,and genetic association at polymorphism loci[J].Cell Research,15(2):97-98.
[26] Vella K R,Burnside A S,Brennan K M,et al.2007.Expression of the hypothalamic transcription factor Nhlh2 is dependent on energy availability[J].Journal of Neuroendocrinology,19(7):499-510.
[27] Xu N,Chen C Y,Shyu A B.1997.Modulation of the fate of cytoplasmic mRNA by AU-rich elements:Key sequence features controlling mRNA deadenylation and decay[J].Molecular and Cellular Biology,17(8):4611-4621.