滩羊TCHH基因CDS区遗传结构及表达模式分析

doi:10.16431/j.cnki.1671-7236.2019.06.003

Abstract

Abstract:

The aim of this experiment was to analyze the genetic structure of TCHH gene in Tan sheep and test the expression pattern of the gene in different tissues of the same month and skin tissue of different months.The full length of the coding sequence (CDS) region of TCHH gene was obtained by homologous cloning of genes,and the homology of the sequence,the basic properties of the protein and the characteristics of the repeat sequence were analyzed by bioinformatics technology.At the same time,the expression pattern of the gene in skin tissue was analyzed by semi-quantitative RT-PCR and Real-time PCR.The results showed that the TCHH gene CDS region was 4 425 bp in length and contained two repeats,encoding a total of 1 474 amino acids.The TCHH amino acid sequence of Tan sheep was highly homologous within the species,with 96.3% homology with Capra hircus,and the homology between species was poor.The homology with Homo sapiens and Mus musculus was 48.4% and 40.5%,respectively.The molecular formula of TCHH protein was C₈₀₀₆H₁₃₁₈₇N₂₇₈₇O₂₆₆₁S₆,the molecular mass was 191.26 ku,and the theoretical isoelectric point was 5.76,which was a hydrophilic protein.Its protein instability index was 119.37,and its stability was poor.The amino acid of this protein contained two conserved regions,S-100 calcium binding domain and protein kinase domain,which were rich in α-helix,up to 89.89%.It was speculated that the secondary structure was a long rod-like molecular structure.The results of tissue expression profiling showed that the gene was specifically expressed in skin tissue,and its mRNA expression decreased with the increase of the age of Tan sheep,which was consistent with the growth trend of the hair curling trait of Tan sheep wool.The specific structure and expression pattern of the TCHH gene indicated that this gene played an important role in the hair curling trait of Tan sheep.This study would provide important reference for the study of the biological mechanism of the occurrence of Tan sheep wool.

Key words: Tan sheep; TCHHgene; expression pattern; curly fleece

CLC Number:

S826

ZHENG Jie, LIU Yufang, MA Lu, XUE Ming. Genetic Structure and Expression Pattern Analysis of TCHH in Tan Sheep[J]. , 2019, 46(6): 1585-1593.

References

[1] 武广善.甘肃滩羊产业现状及发展意见[J].甘肃畜牧兽医,2013,7:11-14. WU G S.Current status and development suggestions of Gansu Tan sheep industry[J].Gansu Animal Husbandry and Veterinary,2013,7:11-14.(in Chinese)
[2] 杨佐青,陶金忠,陈信,等.一周岁内滩羊皮肤差异表达蛋白研究[J].中国畜牧兽医,2017,44(4):986-993. YANG Z Q,TAO J Z,CHEN X,et al.Study on skin differential expression proteins of Tan sheep within one year old[J].China Animal Husbandry & Veterinary Medicine,2017,44(4):986-993.(in Chinese)
[3] LIU Y,KANG X,YANG W,et al.Differential expression of KRT83 regulated by the transcript factor CAP1 in Chinese Tan sheep[J].Gene,2017,614:15-20.
[4] LIU Y,ZHANG J,XU Q,et al.Integrated miRNA-mRNA analysis reveals regulatory pathways underlying the curly fleece trait in Chinese Tan sheep[J].BMC Genomics,2018,19(1):360.
[5] ROTHNAGEL J A,ROGERS G E.Trichohyalin,an intermediate filament-associated protein of the hair follicle[J].Journal of Cell Biology,1986,102(4):1419-1429.
[6] STEINERT P M,PARRY D A D,MAREKOV L N.Trichohyalin mechanically strengthens the hair follicle multiple cross-bridging roles in the inner root sheath[J].Journal of Biological Chemistry,2003,278(42):41409-41419.
[7] LEE S C,LEE J B,SEO J J,et al.Expression of trichohyalin in dermatological disorders:A comparative study with involucrin and filaggrin by immunohistochemical staining[J].Acta Dermato-Venereologica,1999,79(2):122-126.
[8] V BASMANAV F B,CAU L,TAFAZZOLI A,et al.Mutations in three genes encoding proteins involved in hair shaft formation cause uncombable hair syndrome[J].American Journal of Human Genetics,2016,99(6):1292-1304.
[9] WINGET J,FINLAY D,MILLS K J,et al.Quantitative proteomic analysis of stratum corneum dysfunction in adult chronic atopic dermatitis[J].Journal of Investigative Dermatology,2016,136(8):1732-1735.
[10] PETROVA A,CAPALBO A,JACQUET L,et al.Induced pluripotent stem cell differentiation and 3D tissue formation attenuates clonal epigenetic differences in trichohyalin[J]. Stem Cells & Development,2016,25(18):1366-1375.
[11] MEDLAND S E,NYHOLT D R,PAINTER J N,et al.Common variants in the trichohyalin gene are associated with straight hair in Europeans[J].American Journal of Human Genetics,2009,85(5):750-755.
[12] WU S,TAN J,YANG Y,et al.Genome-wide scans reveal variants at EDAR,predominantly affecting hair straightness in Han Chinese and Uyghur populations[J].Human Genetics,2016,135(11):1279-1286..
[13] LIU F,CHEN Y,ZHU G,et al.Meta-analysis of genome-wide association studies identifies 8 novel loci involved in shape variation of human head hair[J].Human Molecular Genetics,2018,27(3):559-575.
[14] ERIKSSON N,MACPHERSON J M,TUNG J Y,et al.Web-based,participant-driven studies yield novel genetic associations for common traits[J].PLoS Genetics,2010,6(6):e1000993.
[15] LEE S C,KIM I G,MAREKOV L N,et al.The structure of human trichohyalin.Potential multiple roles as a functional EF-hand-like calcium-binding protein,a cornified cell envelope precursor,and an intermediate filament-associated (cross-linking) protein[J].Journal of Biological Chemistry,1993,268(16):12164-12176.
[16] TAKASE T,HIRAI Y.Identification of the C-terminal tail domain of AHF/trichohyalin as the critical site for modulation of the keratin filamentous meshwork in the keratinocyte[J].Journal of Dermatological Science,2012,65(2):141-148.
[17] YU Z,GORDON S W,NIXON A J,et al.Expression patterns of keratin intermediate filament and keratin associated protein genes in wool follicles[J].Differentiation,2009,77(3):307-316.
[18] 路立里,狄江,王琼,等.羊毛及动物毛发弯曲形成的生物学机理研究进展[J].畜牧兽医学报,2014,45(5):679-685. LU L L,DI J,WANG Q,et al.Research advances on the biological formation mechanism of wool and hair crimps[J]. Acta Veterinaria et Zootechnica Sinica,2014,45(5):679-685.(in Chinese)
[19] ROGERS G E.Electron microscopy of wool[J].Journal of Ultrastructure Research,1959,2(3):309-330.
[20] MATSUNAGA R,ABE R,ISHⅡ D,et al.Bidirectional binding property of high glycine-tyrosine keratin-associated protein contributes to the mechanical strength and shape of hair[J].Journal of Structural Biology,2013,183(3):484-494.
[21] KIKKAWA Y,OYAMA A,ISHⅡ R,et al.A small deletion hotspot in the type Ⅱ keratin gene mK6irs1/Krt2-6g on mouse chromosome 15,a candidate for causing the wavy hair of the caracul (Ca) mutation[J].Genetics,2003,165(2):721-733.
[22] SCHLAKE T.Krox20,a novel candidate for the regulatory hierarchy that controls hair shaft bending[J].Mechanisms of Development,2006,123(8):641-648.
[23] 刘宁,曹靖,卫盼莹,等.EDAR参与绵羊毛发性状与生长的调节[J].中国生物化学与分子生物学报,2016,32(2):216-223. LIU N,CAO J,WEI P Y,et al.EDAR is involved in the regulation of sheep hair characters and growth[J].Chinese Journal of Biochemistry & Molecular Biology,2016,32(2):216-223.(in Chinese)
[24] KANG X,LIU G,LIU Y,et al.Transcriptome profile at different physiological stages reveals potential mode for curly fleece in Chinese Tan sheep[J].PLoS One,2013,8(8):e71763.
[25] LEE S C,WANG M,MCBRIBE O W,et al.Human trichohyalin gene is clustered with the genes for other epidermal structural proteins and calcium-binding proteins at chromosomal locus 1q21[J].Journal of Investigative Dermatology,1993,100(1):65-68.
[26] ISHIDAYAMAMOTO A,HASHIMOTO Y,MANABE M,et al.Distinctive expression of filaggrin and trichohyalin during various pathways of epithelial differentiation[J].British Journal of Dermatology,1997,137(1):9-16.
[27] STEINERT P M.Biochemical evidence that small proline-rich proteins and trichohyalin function in epithelia by modulation of the biomechanical properties of their cornified cell envelopes[J].Journal of Biological Chemistry,1998,273(19):11758-11769.

Genetic Structure and Expression Pattern Analysis of TCHH in Tan Sheep

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	ZHANG Xueping, SHI Bingang, JIN Xiayang, WANG Xiangyan, LAN Lijuan, SHI Yu, QI Youpeng, ZHAO Shijie, LI Shaobin, HU Jiang. Expression Characterization Analysis of KLF7 Gene and Effects of Its Overexpression on Proliferation and Differentiation of Preadipocytes in Tibetan Sheep [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(8): 2920-2930.
[2]	LI Mingming, WU Qingwei, DENG Yuting, JIN Shengwei, HE Na, ZHANG Junxia. Correlation Analysis of BMPRⅡ Gene Polymorphism and Its Haplotype with Lambing Traits in Tibetan Sheep [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(2): 587-597.
[3]	TIAN Yuan, LONG Feng, LI Anqi, CHEN Jiayue, YANG Sen, ZAN Linsen, CHENG Gong. Amplification, Sequence Feature and Expression Pattern of Snail2 Gene in Qinchuan Cattle [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(9): 3147-3157.
[4]	MA Sijia, LIU Yuan, WANG Xuzhong, LI Tingting, DUAN Xing, YANG Songbai, SONG Dan, LI Xiangchen. Effects of Melatonin on the Expression of Inflammatory Factors in Skeletal Muscle Satellite Cells Stimulated by LPS in Tan Sheep [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(9): 3378-3386.
[5]	GUO Yan, DING Wei, TAO Jinzhong, CHEN Shaoyu, YUAN Ligang, LI Yingkang. Comparison of Skin Histochemical Characteristics in Tan Sheep at Different Ages [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(8): 2762-2770.
[6]	YANG Yuanyuan, HAN Yunsheng, LI Jing, JIA Xueting, ZHAO Qingyu, TANG Chaohua, MA Qing, MA Lina, ZHANG Junmin. Effects of Concentrates and Pasture Feeding on Muscle Fiber Characteristics of Biceps Femoris in Tan Sheep and Its Protein Degradation During Postmortem Aging [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(8): 2797-2808.
[7]	HUO Miao, LI Yanan, YANG Sirui, ZHU Xiangde, XU Xiaofeng, ZHANG Lili. Effect of Feeding Methods on Fatty Acid Composition of Muscle and Adipose Tissue in Tan Sheep Lamb [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(6): 2020-2032.
[8]	ZHANG Hui, LI Xiaoyu, DI Ran, LIU Yufang, CHU Mingxing. Analysis on Difference of Transcription and DNA Methylation Level of AANAT Gene in Ovary of Tan Sheep at Different Reproductive Stages [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(1): 226-234.
[9]	ZHANG Hui, ZHANG Qingyang, LI Chenchen, YANG Junqi, DU Yunliang, LI Kaiyang, LIU Yufang. Cloning and Bioinformatics Analysis of CYP19A1 Gene in Taihang Chickens [J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(5): 1489-1497.
[10]	HUANG Jinxing, CHEN Lan, ZHANG Tao, CHEN Fuxiang, WU Pengfei, MAO Jing, GONG Yongshuang, XIE Kaizhou, WANG Jinyu, DAI Guojun. A Expression Analysis and Target Gene Prediction of miR-206 in Jinmao Black Chickens [J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(2): 506-513.
[11]	WANG Cailian, WU Jianping, LIU Lishan, LANG Xia. Effects of Concentrate Supplementation on Blood Biochemical Indexes in Tibetan Sheep after Grazing at High Altitude Area in Cold Season [J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(12): 3933-3943.
[12]	MU Songyin, XIONG Xianrong, MA Hongcheng, HAI Zhuo, LI Jian. Cloning of L-PGDS Gene and Its Expression Pattern in Different Tissues and Testis at Different Development Stages in Cattle-yak [J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(12): 3985-3992.
[13]	GUO Xiaohong, LIU Cuicui, LI Meng, ZHANG Wanfeng, QIN Benyuan, LIU Min, CAI Chunbo, GAO Pengfei, CAO Guoqing, LI Bugao. Expression Profile and Patterns of CREB-H Gene in Different Tissues and Liver of Pig [J]. China Animal Husbandry and Veterinary Medicine, 2019, 46(9): 2543-2550.
[14]	LI Xianglong, TAO Jinzhong, DING Wei, GUO Yansheng, LIU Guolin, CHEN Yonghong, LIU Huanhuan. Analysis of Wool Traits and Its Relationship in Tan Sheep During the Er-mao Period [J]. , 2019, 46(2): 442-448.
[15]	LI Peiwei, WU Sen, WANG Yaning, WANG Hongbao, ZAN Linsen. Expression Patterns of PLIN2 Gene in Subcutaneous Adipose Tissue and Association Analysis of Single Nucleotide Polymorphisms with Meat Quality Traits in Qinchuan Cattle [J]. , 2018, 45(6): 1580-1589.